climb a ladder是什么意思

Chaos isn"t a pit, Chaos is a ladder. Many who try to climb it fail and never get to try again. The fall breaks them. And some are given a chance to climb, But they refuse. They cling to the realm or the gods or love. Illusions. Only the ladder is real. The climb is all there is. 混乱不是深渊。 混乱是阶梯。 很多人想往上爬 却失败了 且永无机会再试。 他们坠落而亡。 有人本有机会攀爬， 但他们拒绝了。 他们守着王国不放 守着诸神 守着爱情 尽皆幻想。 唯有阶梯真是存在。 攀爬才是生活的全部。

乱世出英雄

《Game of Thrones》作为一部最火美剧，哈佛大学为它专门开了一门—— “The Real Game of Thrones: From Modern Myths to Medieval Models” （真实的权力的游戏：从现代神话到中世纪模型）的课程，课程以改编电视剧为范例，向学生们讲解真实的中世纪。 而加州大学伯克利分校也开了几门与此有关的课程，比如说跟电影研究有关的 “Film Genre: Game of Thrones” （电影类型：权力的游戏）课程，以及 “The Linguistics of Game of Thrones and the Art of Language Invention” 《权力的游戏》语言学及语言创作的艺术”。 走入“冰与火之歌” 这些台词简直是艺术 1、凛冬将至。 Winter is coming. “凛冬将至”，用我们的中文理解便是“居安思危”，在权力的游戏里面没有永远的赢家，各大家族都要懂得一个生存法则——居安思危，时刻提高警惕！ 2、北境不忘。 The North remembers. 3、杀死心中的男孩，真正成长起来。（浴火涅槃） Kill the boy,and let the man be born. 4、你读那么多书有什么用？ Why do you read so much ? 提利昂:我哥哥有他的宝剑，而我有自己的智慧。智慧需要好书滋养，就像宝剑需要磨刀石。 Tyrion:My brother has his sword, and I have my mind. And a mind needs books like a sword needs a whetstone. 5、我试着广交天下好友，你永远不知道哪天谁会派上用场。 I try to know as many people as I can，you never know which one you will need. 6、雄狮不会在乎羊群的想法。 A lion doesn"t concern himself with the opinion of a sheep. 7、混乱不是深渊，混乱是阶梯。 Chaos isn"t a pit. Chaos is a ladder. 8、仇恨是个好东西，让人浑身是劲。 Hate"s as good thing as any to keep a person going. 9、有时候什么都不做才是最难的。 Sometimes nothing is the hardest thing to do. 10、认清自己的长处，明智的加以利用, 一人可对抗万马千军。 Know your strengths, use them wisely, and one man can be worth 10000. 11、长城无懦夫立足之地。 The wall"s no place for cowards. 12、伤痛即是教训，而教训成就未来。 Every hurt is a lesson and every lesson makes you better. 13、物以类聚。 Fools love a fool. 14、兰尼斯特，有债必偿。 A Lannister always pays his debts. 15、没人能保护自取灭亡的人。 No man could protected him from himself. 16、有时候，越有权的人越残忍。 Sometimes those with the most power have least grace. 17、你以为从贫困通往财富的道路永远是纯洁和荣耀的吗？ Do you think the path from poverty to wealth is always pure and honorable？ 18、为得到我想要的东西，我愿意铤而走险。 I"d risk everything to get. what I want. 19、你在乎的人越多，你就越脆弱。 The more people you love, the weaker you are. 20、喝过酒后，任何故事都会更有味道。 There"s no story so good a drink won"t make it better. 21、每个人心中都有一头野兽。 There is a beast in every man. 22、我小心选择同盟，但更谨慎选择敌人。 I choose my allies carefully and my enemies more carefully still. 23、无论我们想要的是什么，一旦得到，就想要更多。 It doesn"t matter what we want, once we get it, then we want something else. 24、有太多人不敢去冒险，一辈子都在躲避危险，结果他们都没能幸免。 So many men, they risk so little, they spend their lives avoiding danger. and then they die. 25、男人们从不渴望他们已经拥有的东西。 Men never crave what they already have. 26、在现实和永远不之间有无限的可能。 A lot can happen between now and never. 27、向自己坦诚真正的自己并不容易。 It" not an easy thing admitting to yourself what you really are. 28、爱是责任的死穴。 love is the death of duty. 29、不管怎样，不要绝望。 Anyway，don"t despair.

风靡世界是肯定有其道理的，权利的游戏：每一句台词都很经典，拿去装逼吧！！Mira Zaslove"The man who passes thesentence should swing the sword." - Eddard Stark.宣判死刑的人，应该亲自挥刀。——奈德 · 史塔克Don"tshy away from making tough calls. And just as importantly, dotheunpleasant work to follow through. As Ned (Eddard Stark) remindsus,"He who hides behind executioners soon forgets what deathis."Leaders who spend time in the trenches, doing the tough work,will takemaking tough decisions more seriously.不要逃避两难的抉择，同样重要的是勇于承担沉重的后果。就像奈德说的那样：“那些躲在刽子手后面的人很快会忘记死亡意味着什么。” 而那些站在第一线、体验严酷工作的领导者会对每一项艰难的决定都非常慎重。"A Lannister always pays hisdebts." - Tyrion Lannister兰尼斯特家族一诺千金。——提利昂 · 兰尼斯特In the workplace, the quickest way to lose respect,and power, is to promisethings you can"t deliver. The surest way to getpeople to do things foryou today, is for them to trust, that you will dowhat you say you will,in the future. Leaders follow through on theirword. When they say they aregoing to do something, they do it.出来混，言而无信会让你迅速失去他人的尊重和你原有的名望。如果你想让别人帮你做事，最保险的方法就是让他人相信你会兑现诺言。一个成功的领导者必须一诺千金。"Any man who must say, I amthe king, is no true king." – Tywin Lannister任何把 "我是国王" 挂在嘴边的人，都不是真正的国王。——泰温 · 兰尼斯特Truepower comes from where people believe it comes from. Not from whereyousay it comes from. The best leaders are followed based on thecollective will,not because they say, “I am the boss.” Power andinfluence, often come fromunexpected places.真正的国王，是人民选的，而不是自己封的。人们跟随一位好的领导者，是出于自愿，而不是因为他说自己是 “老大”。权利和影响力，经常来自意想不到的地方。"Chaos isn"t a pit. Chaos isa ladder." - Little finger混乱的局面下，有些人会掉下去，有些人在向上爬。——小指头Chaotictimes reveal a leader"s strength. When times are good, it"s easy to betheleader. Only when chaos reigns, do many leaders rise. Effectiveleadersaren"t thwarted by challenges. They use challenges to foist themhigher.As Little finger, highlights: "Many who try to climb fail andnever get to tryagain. The fall breaks them.” Leaders are not broken.They continue to climb.混乱的情况下才见一个人的领导力。和平无事的时候，谁都能做好领导。当出现混乱的局面，真正的领导者才会脱颖而出。一个雷厉风行的领导者不会被困难拖住，他们反而会利用这些机会，爬得更高。就像小指头说的：“很多人失败一次就会永远爬不起来。” 而真正的领导者不会被打败，还在继续爬。"Winter is coming." –House Stark寒冬将至——史塔克家族eadersremain vigilant. The world is uncertain. The best leaders alwaysinnovate,stay strong, and plan for the future. Being prepared for theunexpected isessential. Embrace winter, especially when everyone else isdistracted andbasking in the sun.领导者要时刻保持危机感。这个世界充满不确定因素。最好的领导者永远在创新、计划，保持最好的状态，随时准备，拥抱寒冬，特别是当其他人还在享受温暖阳光的时候。《权力的游戏》第七季将来本月与大家见面，一起期待吧。

黑夜漫漫，人心险恶

ladder的读音是：英["l_d_(r)]。ladder的读音是：英["l_d_(r)]。ladder的详尽释义是n.(名词)阶梯，梯子，梯梯状物途径成功的手段梯形裂缝阶层，层次斗架滑丝，（袜子）抽丝进身的阶梯。ladder的词语用法是n.(名词)ladder的基本意思是“梯子”,指便于人上下用的用具,一般用两根长的硬条状物体并排做帮,中间横穿若干根短的横梁制成,是可数名词。引申可表示“阶梯,途径”,是不可数名词。一、详尽释义点此查看ladder的详细内容n.(名词)阶梯，梯子，梯梯状物途径成功的手段梯形裂缝阶层，层次斗架滑丝，（袜子）抽丝进身的阶梯v.(动词)抽丝，袜子抽丝发迹在...上装设梯子，在...架设梯子用梯子爬成名，走红使（袜子）发生抽丝现象，出现滑丝二、英英释义Noun:stepsconsistingoftwoparallelmembersconnectedbyrungs;forclimbingupordownascendingstagesbywhichsomebodyorsomethingcanprogress;"heclimbedthecareerladder"arowofunravelledstitches;"shegotaruninherstocking"Verb:comeunraveledorundoneasifbysnagging;"Hernylonswererunning"三、词典解释1.梯子Aladderisapieceofequipmentusedforclimbingupsomethingordownfromsomething.Itconsistsoftwolongpiecesofwood,metal,orropewithstepsfixedbetweenthem.ladder什么意思2.（社会、组织或系统中晋升或降级的）梯级，阶梯Youcanusetheladdertorefertosomethingsuchasasociety,organization,orsystemwhichhasdifferentlevelsthatpeoplecanprogressupordropdown.e.g.Iftheywanttoclimbtheladderofsuccesstheyshouldbegiventhatopportunity...假如他们有出人头地的意愿，就应该给他们那样的机会。e.g.Sheadmiredhermother"ssisterformovingupthesocialladder.她羡慕姨妈社会地位的提升。3.（女性长筒袜或裤袜的）抽丝，脱线Aladderisaholeortornpartinawoman"sstockingortights,wheresomeoftheverticalthreadshavebroken,leavingonlythehorizontalthreads.e.g.Herhairwasamessandtherewasaladderinhertights.她头发乱糟糟，裤袜脱了线。inAM,use美国英语用run四、例句Diligenceisoftenaladdertosuccess.勤勉常常是通往成功的阶梯。Hewashalfwaydowntheladderwhenhefell.他下阶梯时，半路摔倒了。Thisladderisn"tverystable.这架梯子不太牢靠。It"sconsideredunluckytowalkunderaladder.从梯子下面走过被认为是不吉利的。五、常见句型用作名词(n.)You"llfindFatherontopoftheladder,measuringup.你会看到父亲站在梯子的最高处测量呢。JustwhenIneededtheladder,Ifoundthatyouhadlentittoourneighbour.我正需要用梯子的时候,发现你已经借给了邻居。HandthepaintbrushuptoFatherontheladder.父亲在梯子上,把油漆刷子递给他。Samgotdownfromtheladder.萨姆从梯子上下来了。Getuptheladder,andI"llpassthepaintup.把梯子竖起来,我要把油漆递上去。Wegotuptheladdertotheroof.我们用梯子登上房顶。Iwouldn"tclimbtothetopofthatladder,notevenifyoupaidme.无论如何我也不愿爬到那个梯子的顶端。You"renotsafeonthatladder,abranchcouldknockyouoff.你站在那个梯子上不安全,树枝子会把你碰下来的。Climbtothetopoftheladderandchucktheapplesdowntome.爬到梯子上端,把苹果摘下来扔给我。Cometothebottomoftheladderandchuckuptheotherbrush,willyou?爬下梯子,把另一把刷子扔上来,好吗?Leanaladderagainstawall.把梯子靠在墙上。Wemustgetaladdertogetattheapples.我们得弄个梯子来,才能够着那些苹果。JohnboastedhecouldclimbtotheroofwithoutaladderandsoJimcalledhisbluffandsaid,“Well,let"sseeyoudoit!”约翰吹嘘他不用梯子就能爬上房顶,吉姆逗他说,“好,你爬给我看看。”Heclimbedthesocialladder.他爬上了上层社会。Theonlyladdertoassuresuccessistoworkhard.确保成功的唯一途径是刻苦努力。六、经典引文Heputuptheladderandclimbedtotheloft.出自：R.K.Narayan七、词语用法n.(名词)ladder的基本意思是“梯子”,指便于人上下用的用具,一般用两根长的硬条状物体并排做帮,中间横穿若干根短的横梁制成,是可数名词。引申可表示“阶梯,途径”,是不可数名词。ladder的相关临近词lade、lad、ladderv、laddery、laddered、ladderway、ladderman、laddering、ladderdog、laddergram、ladderlike、laddertie点此查看更多关于ladder的详细信息

Climbaladder放心采纳，不懂可以再问

那个太多了像混沌，谜团，白色，新月，灿烂，无限，死亡呼吸，我就记住这么几个我都是现做现找的，网上就有。

Artist: M2mSong Title: Not To MeYou sent me up to heavenI was safe and loved thereThen you pushed me down <STRONG class=tx_hit>the <STRONG class=tx_hit>ladderNow you just left me hereBut I"m strong, can"t get hurtThis must be wrong, won"t sink down in the dirtWhat"s with meThis is not happy,this is not happyNot to meNot reality...Not to meNot to meMaybe it was my own faultMaybe I just tripped and fellTo blame an angel tempts me nowBaby please take me up againForgive meI didn"t mean to comprehend and consider, my friendWhat"s with meThis is not happy,this is not happyNot to meNot reality...Not to meNot to meBut I"m strong, can"t get hurtThis must be wrong, won"t sink down in the dirtWhat"s with meThis is not happy,this is not happyNot to meNot reality...Not to meNot to meBrett Anderson - The EmpressDidn"t I try To love her?Didn"t I paint Pictures of her?Didn"t I try To change?I shouldn"t have bothered.Didn"t I buy The ocean for her?Didn"t I wear The colourDidn"t I feel The wind?Didn"t I bleed?She is strangeAnd sorrow.She is Like cherry blossom.She is The Empress.Didn"t I kneel Before her?Didn"t I follow Her orders?Didn"t I clean Her shit?Didn"t I fall?Didn"t I climb <STRONG class=tx_hit>The <STRONG class=tx_hit>ladder?Didn"t I smile For the cameras?Didn"t I feel The wind?Didn"t I bleedShe is strange And sorrow.She is Like cherry blossom.She is The Empress.She is strange And sorrow.With lips Like cherry blossom.She is The Empress.Coldplay - 1.36Stuck in a cornerare monkeys in cagesthat don"t have a numberto fight one anothertry to recoverClimb up <STRONG class=tx_hit>the <STRONG class=tx_hit>ladderlook up and you see birdsblind as each otherhow long can we suffer?we"re as blind as each otherOn the cloud that you"re sittingthere"s one born every minuteso much to discoverI"ve become a believerSisters and brotherswho fight one anotherwill mourn and deceive uswill find us and keep ustake us or leave usHow soon is now? yeahhow long is never?I"m nothing but normalwith something togethercome on stick togetherOn the cloud that you"re sittingthere"s one born every minuteso much to discoverI"ve become a believerJennifer Paige - SugarcoatedGo on, go on, go onGo on and tell like it isDon"t want it sugarcoatedOh, your kind of sweet leaves a bad taste in my mouthSo, cmon, cmon, cmonYou never say what"s on your mindIt"s like you"re undercoverYou know how to get your way without making a soundThey"re eating out of the palm of your handsBut they don"t know it"s all a showSugar sweet like honey babySay what they want to hearCuz everybody loves a candied babyMaking it loud and clearWhat"s on the tip of your tongueWhy are you playing dumbGo on just spit it outTake another step aheadAnd climb on up <STRONG class=tx_hit>the <STRONG class=tx_hit>ladderYou"d do anything to get yourself to the topYeah, they"re eating out of the palm of your handsThat"s when you know it"s time to goNever make a stand,take everything you canBruce Springsteen - Working on a DreamOut here the nights are long, the days are lonelyI think of you and I"m working on a dreamI"m working on a dreamNow the cards I"ve drawn"s a rough hand, darlingI straighten the back and I"m working on a dreamI"m working on a dreamI"m working on a dreamThough sometimes it feels so far awayI"m working on a dreamAnd I know it will be mine somedayRain pourin" down, I swing my hammerMy hands are rough from working on a dreamI"m working on a dreamI"m working on a dreamThough trouble can feel like it"s here to stayI"m working on a dreamOur love will chase trouble awayI"m working on a dreamThough it can feel so far awayI"m working on a dreamOur love will make it real somedayThe sun rise up, I climb <STRONG class=tx_hit>the <STRONG class=tx_hit>ladderThe new day breaks and I"m working on a dreamI"m working on a dreamI"m working on a dreamI"m working on a dreamI"m working on a dreamThough it can feel so far awayI"m working on a dreamOur love will make it real somedayI"m working on a dreamThough it can feel so far awayI"m working on a dreamAnd our love will make it real someday本站歌词来自互联网Ah were drinking and were dancingand the band is really happeningand the Johnny Walker wisdom running highAnd my very sweet companionshes the Angel of Compassionshes rubbing half the world against her thighAnd every drinker every dancerlifts a happy face to thank herthe fiddler fiddles something so sublimeall the women tear their blouses offand the men they dance on the polka-dotsand its partner found, its partner lostand its hell to pay when the fiddler stops:its CLOSING TIMEYeah the women tear their blouses offand the men they dance on the polka-dotsand its partner found, its partner lostand its hell to pay when the fiddler stops:its CLOSING TIMEAh were lonely, were romanticand the ciders laced with acidand the Holy Spirits crying, "Wheres the beef?"And the moon is swimming nakedand the summer night is fragrantwith a mighty expectation of reliefSo we struggle and we staggerdown the snakes and up <STRONG class=tx_hit>the <STRONG class=tx_hit>ladderto the tower where the blessed hours chimeand I swear it happened just like this:a sigh, a cry, a hungry kissthe Gates of Love they budged an inchI cant say much has happened sincebut CLOSING TIMEI swear it happened just like this:a sigh, a cry, a hungry kissthe Gates of Love they budged an inchI cant say much has happened sinceCLOSING TIMEI loved you for your beautybut that doesnt make a fool of me:you were in it for your beauty tooand I loved you for your bodytheres a voice that sounds like God to medeclaring, declaring, declaring that your bodys really youAnd I loved you when our love was blessedand I love you now theres nothing leftbut sorrow and a sense of overtimeand I missed you since the place got wreckedAnd I just dont care what happens nextlooks like freedom but it feels like deathits something in between, I guessits CLOSING TIMEYeah I missed you since the place got wreckedBy the winds of change and the weeds of sexlooks like freedom but it feels like deathits something in between, I guessits CLOSING TIMEYeah were drinking and were dancingbut theres nothing really happeningand the place is dead as Heaven on a Saturday nightAnd my very close companiongets me fumbling gets me laughingshes a hundred but shes wearingsomething tightand I lift my glass to the Awful Truthwhich you cant reveal to the Ears of Youthexcept to say it isnt worth a dimeAnd the whole damn place goes crazy twiceand its once for the devil and once for Christbut the Boss dont like these dizzy heightswere busted in the blinding lights,busted in the blinding lightsof CLOSING TIMEThe whole damn place goes crazy twiceand its once for the devil and once for Christbut the Boss dont like these dizzy heightswere busted in the blinding lights,busted in the blinding lightsof CLOSING TIMEOh the women tear their blouses offand the men they dance on the polka-dotsIts CLOSING TIMEAnd its partner found, its partner lostand its hell to pay when the fiddler stopsIts CLOSING TIMEI swear it happened just like this:a sigh, a cry, a hungry kissIts CLOSING TIMEThe Gates of Love they budged an inchI cant say much has happened sinceBut CLOSING TIMEI loved you when our love was blessedI love you now theres nothing leftBut CLOSING TIMEI miss you since the place got wrecked 欢迎您的光临! Just spit it out

西方的迷信，从工作梯下走过。他们认为：If you walk under a ladder, you will have bad luck. 如果从工作梯下走过，会带来坏运气 无论工作梯上有没有人，从下面走过就会带衰．．．但我想，若是有人在上头工作的话，基于安全考虑应该生人都是会回避才是；若是没有人的话，梯子．．．应该会被收起来吧！ (就是从这家伙下面走过会带衰)

我认为楼上的前面三句是可以完全不需要的，因为根本不是这个问题。还有，更正楼上的一个错误，就算你是改成从句的形式，也应该是：I saw a ladder that was leaning against the building.而不是I saw a ladder that is leaning against the building. 应该把is 改成was.还有我认为，I saw a ladder leaning against the building. 直接翻译成“我看到一把梯子斜靠在建筑物上”就行了，不用“正在”那么别扭。其它没什么了，说得很好。

green ladder应该是指剧情中的一个物品（或是比喻）。这就要看前后关联了，green ladder和brother分别是两个暗号，好好看看剧情就应该能知道了。green ladder 没有特殊的含义。

Student: So you"re saying … like, if you had a ladder tall enough, you could just climb into space? 那么你是说，如果有一个足够高的梯子，人们可以爬进太空? 学生用疑问的语气，也就是在问教授自己的理解对不对，因此选择 B 。

ladder is more a 梯子and gangway is more a 通道。。。 查看原帖>>

谓语

A

从梯子下散步

下步行梯。

He knows your name, boyYou"ll write it downUnder bridges into townAnd whats that rusting?, you"re fathers crown!And our mother sleeps in the groundI go out walking, my own feetAnd I might sing, "Oh bury me"And from the bedroom exudes the awful soundAnd our mother sleeps in the groundOh, snakes on the ladderOh, my eyes were something i sawThere"s a motion in the skyAnd the singer who called outsideTo the woods reflecting back a million soundsOf our lives, inside the forest off Greyhound?Oh, Snakes on the ladderOh, my eyes were something i saw

c下来！请采纳，谢谢

B 本题我们极易错选A或C。That"s all 意思是“就这些”，表示讲话或文章的结束，而That"s all right 意思是“行；可以；没关系”。That"s it 为固定搭配，意思是“这正是所需要的”。

这题的正确答案：选C.broad 宽阔的 这个房间是三米长,两米宽. broad 是形容词作表语,和前面的形容词long对应,其它的词都不对 width是名词,board 是名词,板的意思,abroad 是国外的,与题意不符, 所以只有C是正确的.

Let us use your ladder truck . - get up now 让我使用你的云梯车吧起来了！ When a new york fire department ladder truck .纽约消防队的一辆云梯车 We " ve got no business with you . - we need a ladder truck 我们和你没有生意来往我们需要云梯车！ We " ve got a ladder truck ing . have them move their cars 我们有辆云梯车要进来叫他们把车子移一下 We " re waiting for the ladder truck 我们正在等云梯车 Three days later fireman bob picked up billy , dressed him in his fire uniform and escorted him from his hospital bed to the ladder truck 三天后，消防员鲍勃来接比利，为他穿上消防制服，从医院病床一直陪同来到装著长梯的消防车上。 About five minutes later a ladder truck arrived at the hospital , extended its ladder up to billy " s third floor open window and 5 firefighters cpmbed up the ladder into billy " s room 约莫五分钟后，装著长梯的消防车到了医院，将梯子伸到三楼比利敞开的窗户上，五名消防员爬上梯子进入比利的房间。 It"s difficult to find ladder truck in a sentence. 用 ladder truck 造句挺难的

ladder stitchna.梯形(图案)刺绣法阶梯绣；梯形花样的刺绣；梯形刺绣.-----------------------------------为你解答，如有帮助请采纳，如对本题有疑问可追问，Good luck!

生命悬于从直升机放下的绳梯上靠着直升机的绳梯救命PS 翻译是可以很灵活的，不要教条的去翻译。

She fell off a ladder and broke her arm . 她从梯子上摔下来，跌断了胳膊。 The ladder is leaning up against the wall . 那个梯子靠著墻呢。 Have you any tights that wo n"t ladder ? 你有不抽丝的裤袜吗？ I got a ladder from the shed . 我从花园的小棚里拿来一架梯子。 He lost his balance and fell from the ladder .. 他失去平衡，从梯子上摔下来。 Tom set the ladder against the wall . 汤姆把梯子靠在墻上。 The ladder will be more secure here . 梯子靠这儿比较牢稳。 He fell off the ladder and landed bump on the ground . 他从梯子上砰的一声跌到地上。 Nylon tights sometimes ladder . 尼龙裤袜有时候抽丝。 The ladder rested against the wall . 梯子倚在墻上。 It"s difficult to see ladder in a sentence. 用 ladder 造句挺难的 Rest the ladder against the wall . 把梯子靠在墻上。 She laddered her new tights cpmbing the fence . 她爬越篱笆时把她的裤袜给勾得抽了丝。 She got down from the ladder . 她从梯子上下来。 In one corner a ladder went up to an open trap door . 角落里有一道梯子，通往上面敞开的活板门。 Slope the ladder a bit . 把梯子放坡点。 I cpmbed up on a high ladder with a carton of fresh eggs . 我手拿一盒鲜鸡蛋，爬上一把很高的梯子。 But he was about to blow up the ladder he was standing on . 但是他即将会毁掉他现在站立的这个阶梯。 He was a tenacious beast. on the very ladder he pngered . 他这死不甘心的家伙，在舷梯上踌躇不前了。 Is that ladder safe ? 那梯子结实吗？ She was fast asleep , so i got a ladder from the shed . 她睡得很熟，于是我从花园的小棚里拿来一架梯子。 Ladders - part 1 : terms , types , functional sizes 梯子第1部分:术语型式和功能尺寸 Scapng ladder - one who was fearless in attacking 梯子在拼斗中勇往直前的人。 Loading test of acmodation ladder ( pilot ladder ) 舷梯（引水员梯）压重试验。 Scapng ladder - one who was fearless in attacking 梯子在进攻中勇往直前的人。 Hand the paintbrush up to father on the ladder 把漆刷递上去交给梯子上的父亲。 Lower the pilot ladder on the port starboard side 把领航员的包从领航艇上吊上来 We " re gonna need a human ladder starting right . here 我们需要一道人梯从这里开始 It " s considered unlucky to walk under a ladder 从梯子下面走过被认为是不吉利的。 The pttle boy had an unsteady footing on the ladder 那小孩在梯子上站得不稳。 It"s difficult to find ladder in a sentence. 用 ladder 造句挺难的

Climb a ladder放心采纳，不懂可以再问

歌曲名:White Ladder歌手:David Gray专辑:White LadderMusicWhite ladderWater and wineForever cryingOh timeDon"t wanna feel tonightThere"s no rhyme or reason to lifeThis sweet lifeWhite riverThunder and warOh noTher"s no rhyme or reason to lifeThis seet lifeTo the night another bodyTo the night another nameTo the night another valentineA burning flamehttp://music.baidu.com/song/7870533

它10米长 ,宽,高 It is 10 meters long and 10 meters in height.同意 it is 10 meters in length / width / height ..

1、上下楼道靠右行，你谦我让脚步轻。 On the corridor on the right side, I let you modest light footsteps. 2、上楼下楼靠右走，遇到混乱手牵手。 Up and down the stairs on the right hand, encountered confusion. 3、危险地方我不去，学会保护我自己。 I won"t go where I"m in danger, and learn to protect myself. 4、上下楼梯靠右行，安全意识时时有。 Keep to The Right, have a sense of safety. 5、若以追跑为开心，将以后悔而伤心。 If the chase for fun, will regret and sad. 6、楼道走廊不嬉戏，同学玩乐守秩序。 Corridor corridor does not play, students play, keep order. 7、高高兴兴上学来，平平安安回到家。 Come to school cheerfully, and return home safe and sound. 8、上下小心，脚步轻轻。 Watch your step, step softly. 9、上下楼梯不拥挤，集体活动守纪律。 The stairs are not crowded, and the collective activities are disciplined. 10、课间活动要文明，不可追逐和打闹。 Activities should be civilized, not chase and slapstick. 11、前面混乱我看见，不要走过去添乱。 I don"t see the chaos in front of, go up. 12、走楼梯上下楼梯，请靠右行。 Take the stairs, keep right. 13、上下楼梯，请靠右行。 Up and down the stairs, keep right. 14、自己摔倒不要慌，大喊呼救叫帮忙。 Don"t panic when you fall, shout for help and call for help. 15、上下楼梯，相互礼让。 Walk up and down the stairs and show courtesy to each other. 16、教室走廊不嬉戏，同学玩乐守秩序。 Classroom corridors do not play, students play, keep order. 17、人多行走不拥挤，安全永远伴我行。 Many people walk, not crowded, safe with me forever. 18、上下楼梯靠右行，严禁拥挤保安全。 Keep to The Right, crowded security is strictly prohibited. 19、不挤不急，安全第一。 No rush, safety first. 20、维护校园安全，营造和谐环境。 Maintain campus safety and create a harmonious environment. 21、金钱有价，生命无价。 Money is valuable, life is priceless. 22、课间游戏要安全，上下楼梯不打闹。 Break the game to be safe, up and down the stairs, no slapstick. 23、楼梯走廊不嬉戏，同学玩乐守秩序。 Stairs, corridors do not play, students play, keep order. 24、不要推不要挤，关心别人爱自己。 Don"t push, don"t push, care about others, love yourself. 25、上下楼梯靠右走，你谦我让脚步轻。 Up and down the stairs right away, I will let you first light footsteps. 26、不挤不拥不打闹，怀文校园更美好。 Do not squeeze hold not slapstick, Huaiwen campus more beautiful. 27、轻声慢步不打闹，上下楼梯要走好。 Walk softly not slapstick, up and down the stairs to go. 28、楼道走廊不奔跑，安全第一真是好。 Corridor corridor does not run, safety first, really good. 29、上下楼梯慢慢行，楼梯上下不蹦跨。 Go up and down the stairs slowly, the stairs do not jump up and down. 30、同学摔倒楼梯上，自己伸手快帮忙。 When students fall on the stairs, they help themselves. 31、走廊内不奔跑，不追逐不打闹。 Corridors do not run, do not chase, do not fight. 32、轻声细语，举止文明。 Speak softly and behave with civility. 33、行走靠右，谨防摔伤。 Walk on guard against falls. 34、楼梯通道，请勿跑跳。 Don"t run, please. 35、上下楼梯靠右走，安全意识时时有。 Up and down the stairs right away, a sense of safety. 36、课间活动有秩序，校园文明争第一。 During the break, the activities are orderly, and the campus civilization is the first. 37、人生美好，步步小心。 Life is good, step by step, careful. 38、讲究卫生，保持清洁。 Pay attention to hygiene and keep clean. 39、上下楼梯靠右行，千万不能滑下梯。 Keep to The Right, do not slide down the ladder. 40、轻声漫步，同学玩乐守秩序。 Walk softly, students play, keep order. 41、学生就要爱学校，上楼下楼不打闹。 Students should love the school, upstairs, downstairs, not slapstick. 42、安全关系你我他，校园安全靠大家。 Security concerns you, me, him, campus security depends on everyone. 43、老师就像我妈妈，安全知识要听她。 The teacher is like my mother, and the safety knowledge must be heard by her. 44、请轻轻的走路，悄悄的说话。 Please walk softly and speak quietly. 45、幸福是棵树，安全是沃土。 Happiness is a tree, and safety is the soil. 46、快快乐乐上学，平平安安回家。 Go to school happily, go home safe and sound. 47、上下楼梯靠右走，你追我赶危险多。 Up and down the stairs right away, race each other more dangerous. 48、课余时间不疯狂，损坏公物易受伤。 Spare time is not crazy, damage to public property is easy to hurt. 49、轻声慢步，文明上下楼。 Quietly walk down the stairs, civilization. 50、幸灾乐祸笑哈哈，自己出事没人拉。 Gloat, laugh, and no one pulled his own accident. 51、下课之后请慢走，追逐打闹不安全。 After class I"m chasing is not safe. 52、上下楼梯靠右行，轻声慢步讲文明。 Keep to The Right, quietly walk civilization. 53、脚步轻轻，靠右慢行。 Softly, right slow. 54、生命至高无上，安全责任为天。 Life is suPme, security is duty. 55、右行谦让，举止文明。 Right line humility, civilized manners. 56、走廊楼道不跑跳，脚步要轻慢慢行。 Corridor corridor does not run, jump, the pace should be light, slow. 57、下课不疯跑，小心被摔倒。 Don"t run crazy after class, be careful to fall. 58、脚步轻轻，勿扰他人。 Softly and others. 59、楼梯走廊不奔跑，安全第一真是好。 Stair corridor does not run, safety first is really good. 60、上下楼梯不打闹，安全对你很重要。 Safety is important to you when you walk up and down stairs. 61、不要跑，不要跳，不要打闹向右靠。 Do not run, do not jump, do not rough, lean to the right. 62、安全在我心中，生命在我手中。 Safety is in my heart, life is in my hands. 63、伤害别人，等于伤害自己。 He who injures others is equal to himself. 64、玩耍嬉戏要当心，安全警钟常在心。 Be careful when you play and play, and your safety alarm is always on your mind. 65、上上下下，注意脚下。 Up and down, watch your step. 66、上下楼梯能互让，通道自然就宽敞。 The stairs can naturally spacious accommodation, channel. 67、不挤不拥不打闹，文明校园更美好。 No crowded, no fight, civilized campus better. 68、平安是金，步步小心。 Peace is gold, step by step. 69、扔下的是废纸，捡起的是品质。 The waste paper was dropped and the quality was picked up. 70、上下楼梯靠右行，不推不挤步步稳。 Keep to The Right, not able to squeeze. 71、楼梯非跑道，上下勿奔跑。 The stairs are off the track, don"t run up and down. 72、上下楼梯，切勿打闹。 Walk up and down stairs, never fight. 73、轻声慢步，倡导文明好风尚。 Walk softly, advocate civilized morality. 74、结伴同行不打闹，单独行走不抢行。 Go together without a fight, walk alone, do not rob. 75、走楼梯守秩序，进教室不喧哗。 Take the stairs, keep order, and get into the classroom without making noise. 76、安全是朵幸福花，大家浇灌美如画。 Security is a happy flower watering, everyone is. 77、守秩序，进教室不喧哗。 Keep order and enter the classroom without making noise. 78、集体行动列队走，安全两字记在心。 Collective action parade, safe word in mind. 79、上下楼梯靠右走，以免摔倒碰破头。 Up and down the stairs right away, so as not to fall Pengpo head. 80、轻声漫步不打闹，上下楼梯要走好。 Walk softly, don"t fight, walk up and down the stairs. ;

短文填词 1 meaning 2 called 3 bad 4 so 5 (应该是whether把)to think 6 to 7 unmindfully 8 （是surprised him把.）fall 9 down 10 pooled 同义词： feels tired 填空： 1 notice 2 pen

它在爬梯子。

歌曲名:Homeworld [The Ladder]歌手:Yes专辑:The LadderNothing can take us far enough emotionFar enough togetherAs the light shines so brightBright enough to let us inNothing can bridge our souls" devotionFast enough togetherAs the power proves you rightRight enough to let you beginSo many displaced among the future dreamersRealized their doublesTook a new stepA question of originOnly in the recent pastSeekers fought to realizeSkyward shone like beaconsA question of originTen thousand millions fleeTo the westward lightThe dreamers representThis arc of peaceAs the poets entrancedThe anchor redeemedSecrets of scienceThe history of the futureWas surely madeJust what keeps us so aliveJust what makes us realizeOur home is our world, our lifeHome is our worldNothing can take us far enough emotionFar enough togetherAs the light shines so brightBright enough to let us inNothing can bridge our souls devotionFast enough togetherAs the power proves you rightRight enough to let you beginSpeak so fast to the prophets of the livingLooking for the signsSpanning out the centuriesSearch for truthAncient ones, they watch and listenCarry our wishesTook upon themselves to guide usThrough the endless skiesJust what keeps us so aliveJust what makes us realizeOur home is our world, our lifeOur hope is our world, our lifeI have seen the passionThat"s in the hope that everyoneWill find their way into the secretOf the home of your heartLiving within the visionWithin the power, beyond beliefWe see that hate destroys the soulOf anyone who tries to teach itI have seen the dreamThat"s in your heartThat"s in your eyesTo bring you closer to the oneIt"s what keeps us so aliveIt"s what makes us realizeOur home is our world, our lifeJust what keeps us so aliveJust what makes us realizeOur home is our world, our lifeHome is our world, our lifeSend, ascending to the secretsAll is pure and clear to resolveNothing can change us nowSend, ascending to the futureNothing can ever change us nowWe follow the sunWe follow the sunWe follow the sunTruth is a simple placeHere for us all to seeReach as it comes to youAs it comes to meAs I will always need you inside my heartPeace is a word we teachA place for us all to reachSing as it sings to youAs it sings to meAs I will always need you inside my hearthttp://music.baidu.com/song/24535655

英语谚语: He who would climb the ladder must begin at the bottom 中文意思: 爬梯必须从底层开始。 随机推荐10条英文谚语： He who has health has hope and he who has hope everything 身体好就有希望，有希望就有一切。 He who has lost his good name is a dead man among the living 丧失好名誉，犹如活死人。 He who has lost shame is lost to all virtue 没有廉耻心，没有好德行。 He who has no shame has no conscience 没有廉耻的人，没有良心。 He who has not tasted bitter knows not what sweet is 不吃苦中苦，哪知甜中甜。 He who hesitates is lost 举棋不定，坐失良机。 He who imagines that he has knowledge enough has none 自以为学识丰富的人是没有什么学问的人。 He who is afraid of asking is ashamed of learning 害怕问者耻于学。 He who is ashamed of asking is ashamed of learning 耻于问者耻于学。 He who is born a fool is never cured 生成的傻子总是治不好的。 英语谚语: He who would climb the ladder must begin at the bottom 中文意思: 爬梯必须从底层开始。

这是主系表结构加定语从句的用法。courage is the ladder 这是主系表成分。on which all the other virtues mount这是 on which引导的定语从句，修饰前面的ladder。

Jacob"s LadderChumbawambaJacob"s Ladder演唱：Chumbawamba歌曲：Jacob"s Ladder (Not in My Name)~幸魂~收藏like a sermon on the mountainsays the dumber got dumbhellfire and brimstoneswapped for oil and gunswhen we"re pushing up daisieswe all look the samein the name of the father maybebut not in my nameon this Jacob"s ladderthe only way up is downone step from disastertwo to make the higher groundJacob"s ladderAnd they sent him to the wars to be slain, to be slainAnd they sent him to the wars to be slaina million lifetimesleft dying in the sunin the streets down in Whitehalldogs picking at the bonesnine eleven got brandednine eleven got soldthere will be no one left to waterall the seeds you"ve sownon this Jacob"s ladderthe only way up is downone step from disastertwo to make the higher groundJacob"s ladderand they sent him to the warsto be slain to be slainand they sent him to the warsto be slainAnd they sent him to the wars to be slain, to be slainAnd they sent him to the wars to be slainon this Jacob"s ladderthe only way up is downone step from disastertwo to make the higher groundon this Jacob"s ladderthe only way up is downone step from disastertwo to make the higher groundJacob"s ladderO Bulldog Leader,sooner or laterwe"ll dig up the body and try her murderO Bulldog Leader,sooner or laterwe"ll dig up the body and try her murder........http://music.baidu.com/song/8368539

千里之行，始于足下

cubs是不是那只狮子的名字？如果是的话，改成一般疑问句就是：Can the lion Cubs climb up a ladder?如果是你打错的话……就是：Can the lion climb up a ladder?

I got him ! i got him ! - ladder 49 to mand 我找到他了，我找到了-云梯49报告总部 Lenny ) is everybody all right ? - ladder 49 to mand 大家都没事吗？ -云梯49报告 - is everybody all right ? - ladder 49 to mand -大家都没事吗？ -云梯49报告 - i got him ! i got him ! - ladder 49 to mand -我找到他了，我找到了-云梯49报告总部 Lenny is everybody all right ? - ladder 49 to mand -大家都没事吗？ -云梯49报告 Ladder 49 , there " s a control room straight ahead 云梯49 ，正前方有个控制室 Ladder 49 to mand . we " re on the 1 2th floor 49号云梯报告我们在12楼 Ladder 49 to mand . we got a man in " over the top 云梯49报告有个队员从顶层下来了 ( kennedy ) ladder 49 , i need a situation report 云梯49 ，我要进度报告 I need a goddamn progress report , ladder 49 我需要进度报告，云梯49 Kennedy ) ladder 49 , i need a situation report 云梯49 ，我要进度报告 Kennedy ladder 49 , i need a situation report 云梯49 ，我要进度报告 Command to ladder 49 . jack , e in 这里是总部向云梯49杰克，报告 Anybody with him ? - the others of ladder 49 made it out 有人和他在一起吗？ -云梯49的其他人都出来了 - anybody with him ? - the others of ladder 49 made it out -有人和他在一起吗？ -云梯49的其他人都出来了 Ladder 49 , morrison to mand 云梯49 ，莫里森向总部 Truck 51 to ladder 49 portable 51号车向云梯49分队 The film is produced by casey silver ( ladder 49 , hidalgo , the uping leatherheads ) of casey silver productions 电影制作是由凯西银( 49个阶梯,伊达尔戈,即将leatherheads )凯西白银制作

两尖阶梯

Language in mission 与College：条件关系if you pursue certain Language in mission，you have to accomplish by college education.The Ladder to success与Discovery of a new life stage ：因果关系The Discovery of a new life stage is a key step to find out the Ladder to success.so，if you pursue the Ladder to success in your life ，you have to set up your goal such as to pursue certain Languagec in college and by doing so，you first need to adopt a new style of life.of course，there are other logic relationships to interpret those；just make them accountible.

career ladder的意思就是：职务级别提升，（职务）提级。

卖房

FANUC_Fladder_Ⅲ_V5.7汉化补丁双语版.rarhttp://blog.sina.com.cn/s/blog_6a00c7be0101cfga.html5.7 6.3在我的博客也可以找到。

梯田 梯形 梯队 阶梯。。云梯。。。。电梯。。。。木梯。。。

你并不孤单 因为我始终在你身旁 尽管你已走远 尽管我还在这里 你并不孤单 因为我始终在你身旁 尽管我们相隔万水千山 你却时刻在我心中

更新1: 我都知系状态...但系有分别嫁.... 我指紧系science..... phys chem....一定有分别．．．．． 无分别就唔会有两个字啦．．．． phase同state都有同样既意思 -- 状态 但系science通常会用phase 因为佢同时有阶段既意思 至于state有好多解释(状态只系其中一个) 例如:国家 州 情况等 You can say in Science World phase = period state = performance

先给你贴个东西你先看一下 希望对你有帮助 将特定的外源基因构建在植物表达载体中并转入受体植物,并不是植物遗传转化的最终目的.理想的转基因植物往往需要外源基因在特定部位和特定时间内高水平表达,产生人们期望的表型性状.然而,近二十年的发展历史却表明,外源基因在受体植物内往往会出现表达效率低、表达产物不稳定甚至基因失活或沉默等不良现象,导致转基因植物无法投入实际应用.另外,转基因植物的安全性问题已在许多国家引起人们的关注,例如,转基因有可能随花粉扩散,抗生素筛选标记基因有可能使临床上的某些抗生素失去作用等等.以上问题的出现使得植物基因工程这一高新技术正处于一种前所未有的困扰时期.针对这些问题,近几年人们对植物转基因技术进行了多方面的探索和改进,植物表达载体的改进和优化就是其中最重要的一项内容,本文就已经取得的进展进行综述. 1 启动子的选用和改造 外源基因表达量不足往往是得不到理想的转基因植物的重要原因.由于启动子在决定基因表达方面起关键作用,因此,选择合适的植物启动子和改进其活性是增强外源基因表达首先要考虑的问题. 目前在植物表达载体中广泛应用的启动子是组成型启动子,例如,绝大多数双子叶转基因植物均使用CaMV35S启动子,单子叶转基因植物主要使用来自玉米的Ubiquitin启动子和来自水稻的Actinl启动子.在这些组成型表达启动子的控制下,外源基因在转基因植物的所有部位和所有的发育阶段都会表达.然而,外源基因在受体植物内持续、高效的表达不但造成浪费,往往还会引起植物的形态发生改变,影响植物的生长发育.为了使外源基因在植物体内有效发挥作用,同时又可减少对植物的不利影响,目前人们对特异表达启动子的研究和应用越来越重视.已发现的特异性启动子主要包括器官特异性启动子和诱导特异性启动子.例如,种子特异性启动子、果实特异性启动子、叶肉细胞特异性启动子、根特异性启动子、损伤诱导特异性启动子、化学诱导特异性启动子、光诱导特异性启动子、热激诱导特异性启动子等.这些特异性启动子的克隆和应用为在植物中特异性地表达外源基因奠定了基础.例如,瑞士CIBA-GEIGY公司使用PR-IA启动子控制转基因烟草中Bt毒蛋白基因的表达,由于该启动子可受水杨酸及其衍生物诱导,通过喷酒廉价、无公害的化学物质,诱导抗虫基因在虫害重发生季节表达,显然是一个十分有效的途径. 在植物转基因研究中,使用天然的启动子往往不能取得令人满意的结果,尤其是在进行特异表达和诱导表达时,表达水平大多不够理想.对现有启动子进行改造,构建复合式启动子将是十分重要的途径.例如,Ni等人将章鱼碱合成酶基因启动子的转录激活区与甘露碱合成酶基因启动子构成了复合启动子,GUS表达结果表示：改造后的启动子活性比35S启动子明显提高.吴瑞等人将操作诱导型的PI-II基因启动子与水稻Actinl基因内含子1进行组合,新型启动子的表达活性提高了近10倍（专利）.在植物基因工程研究中,这些人工组建的启动子发挥了重要作用. 2 增强翻译效率 为了增强外源基因的翻译效率,构建载体时一般要对基因进行修饰,主要考虑三方面内容： 2.1添加5‘-3‘-非翻译序列 许多实验已经发现,真核基因的5‘-3‘-非翻译序列（UTR）对基因的正常表达是非常必要的,该区段的缺失常会导致mRNA的稳定性和翻译水平显著下降.例如,在烟草花叶病毒（TMV）的126kDa蛋白基因翻译起始位点上游,有一个由68bp核苷酸组成的Ω元件,这一元件为核糖体提供了新的结合位点,能使Gus基因的翻译活性提高数十倍.目前已有许多载体中外源基因的5‘-端添加了Ω翻译增强序列.Ingelbrecht等曾对多种基因的 3‘-端序列进行过研究,发现章鱼碱合成酶基因的3‘-端序列能使NPTII基因的瞬间表达提高20倍以上.另外,不同基因的3‘-端序列增进基因表达的效率有所不同,例如,rbcS3‘-端序列对基因表达的促进作用比查尔酮合酶基因的3‘-端序列高60倍. 2.2 优化起始密码周边序列 虽然起始密码子在生物界是通用的,然而,从不同生物来源的基因各有其特殊的起始密码周边序列.例如,植物起始密码子周边序列的典型特征是AACCAUGC,动物起始密码子周边序列为CACCAUG,原核生物的则与二者差别较大.Kozak详细研究过起始密码子ATG周边碱基定点突变后对转录和翻译所造成的影响,并总结出在真核生物中,起始密码子周边序列为ACCATGG时转录和翻译效率最高,特别是-3位的A对翻译效率非常重要.该序列被后人称为Kozak序列,并被应用于表达载体的构建中.例如,有一个细菌的几丁质酶基因,原来的起始密码周边序列为UUUAUGG,当被修饰为ACCAUGG,其在烟草中的表达水平提高了8倍.因此,利用非植物来源的基因构建表达载体时,应根据植物起始密码子周边序列的特征加以修饰改造. 2.3对基因编码区加以改造 如果外源基因是来自于原核生物,由于表达机制的差异,这些基因在植物体内往往表达水平很低,例如,来自于苏云金芽孢杆菌的野生型杀虫蛋白基因在植物中的表达量非常低,研究发现这是由于原核基因与植物基因的差异造成了mRNA稳定性下降.美国Monsanto公司Perlak等人在不改变毒蛋白氨基酸序列的前提下,对杀虫蛋白基因进行了改造,选用植物偏爱的密码子,增加了GC含量,去除原序列下影响mRNA稳定的元件,结果在转基因植株中毒蛋白的表达量增加了30～100倍,获得了明显的抗虫效果. 3 消除位置效应 当外源基因被移人受体植物中之后,它在不同的转基因植株中的表达水平往往有很大差异.这主要是由于外源基因在受体植物的基因组内插入位点不同造成的.这就是所谓的"位置效应".为了消除位置效应,使外源基因都能够整合在植物基因组的转录活跃区,在目前的表达载体构建策略中通常会考虑到核基质结合区以及定点整合技术的应用. 核基质结合区（matrix association region,MAR）是存在于真核细胞染色质中的一段与核基质特异结合的DNA序列.一般认为,MAR序列位于转录活跃的DNA环状结构哉的边界,其功能是造成一种分割作用,使每个转录单元保持相对的独立性,免受周围染色质的影响.有关研究表明,将MAR置于目的基因的两侧,构建成包含MAR-gene-MAR结构的植物表达载体,用于遗传转化,能明显提高目的基因的表达水平,降低不同转基因植株之间目的基因表达水平的差异,减少位置效应.例如,Allen等人研究了异源MAR（来自酵母）和同源MAR（来自烟草）对Gus基因在烟草中表达的影响,发现酵母的MAR能使转基因表达水平平均提高12倍,而烟草本身的MAR能使转基因的表达水平平均提高60倍.使用来源于鸡溶菌酶基因的MAR也可起到同样作用. 另一可行的途径是采用定点整合技术,这一技术的主要原理是,当转化载体含有与寄主染色体同源的DNA片段时,外源基因可以通过同源重组定点整合于染色体的特定部位.实际操作时首先要分离染色体转录活性区域的DNA片段,然后构建植物表达载体.在微生物的遗传操作中,同源重组定点整合已成为一项常规技术,在动物中外源基因的定点整合已获得成功,而在植物中除了叶绿体表达载体可实现定点整合以外,细胞核转化中还很少有成功的报道. 4 构建叶绿体表达载体 为了克服细胞核转化中经常出现的外源基因表达效率低,位置效应及由于核基因随花粉扩散而带来的不安全性等问题,近几年出现的一种新兴的遗传转化技术--叶绿体转化,正以它的优越性和发展前景日益为人们所认识并受到重视.到目前为止,已在烟草、水稻、拟南芥、马铃薯和油菜（侯丙凯等,等发表）5种植物中相继实现了叶绿体转化,使得这一转化技术开始成为植物基因工程中新的生长点. 由于目前多种植物的叶绿体基因组全序列已被测定,这就为外源基因通过同源重组机制定点整合进叶绿体基因组奠定了基础,目前构建的叶绿体表达载体基本上都属于定点整合载体.构建叶绿体表达载体基本上都属于定点事例载体.构建叶绿体表达载体时,一般都在外源基因表达盒的两侧各连接一段叶绿体的DNA序列,称为同源重组片段或定位片段（Targeting fragment）.当载体被导入叶绿体后,通过这两个片段与叶绿体基因组上的相同片段发生同源重组,就可能将外源基因整合到叶绿体基因组的特定位点.在以作物改良为目的的叶绿体转化中,要求同源重组发生以后,外源基因的插入既不引起叶绿体基因原有序列丢失,又不致于破坏插入点处原有基因的功能.为满足这一要求,已有的工作都选用了相邻的两个基因作为同源重组片段,例如rbcL/accD,16StrnV/rpsl2rps7,psbA/trnK,rps7/ndhB.当同源重组发生以后,外源基因定点插入在两个相邻基因的间隔区,保证了原有基因的功能不受影响.最近,Daniel等利用烟草叶绿体基因trnA和trnI作为同源重组片段,构建了一种通用载体（universal vector）.由于trnA和trnI的DNA序列在高等植物中是高度保守的,作者认为这种载体可用于多种不同植物的叶绿体转化.如果这种载体的通用性得到证实,那么这项工作无疑为构建方便而实用的新型叶绿体表达载体提供了一个好的思路. 由于叶绿体基因组的高拷贝性,定点整合进叶绿体基因组的外源基因往往会得到高效率表达,例如McBride等人首次将Bt CryIA(c)毒素基因转入烟草叶绿体,Bt毒素蛋白的表达量高达叶子总蛋白的3%～5%,而通常的核转化技术只能达到0.001%～0.6%.最近,Kota等将Bt Cry2Aa2蛋白基因转入烟草转入烟草叶绿体,也发现毒蛋白在烟草叶子中的表达量很高,占可溶性蛋白的2%～3%,比细胞核转化高出20～30倍,转基因烟草不仅能抗敏感昆虫,而且能够百分之百地杀死那些产生了高抗性的昆虫.Staub等最近报道,将人的生长激素基因转入烟草叶绿体,其表达量竟高达叶片总蛋白的7%,比细胞核转化高出300倍.这些实验充分说明,叶绿体表达载体的构建和转化,是实现外源基因高效表达的重要途径之一. 5 定位信号的应用 上述几种载体优化策略主要目的是提高外源基因的转录和翻译效率,然而,高水平表达的外源蛋白能否在植物细胞内稳定存在以及积累量的多少是植物遗传转化中需要考虑的另一重要问题. 近几年的研究发现,如果某些外源基因连接上适当的定位信号序列,使外源蛋白产生后定向运输到细胞内的特定部位,例如：叶绿体、内质网、液泡等,则可明显提高外源蛋白的稳定性和累积量.这是因为内质网等特定区域为某些外源蛋白提供了一个相对稳定的内环境,有效防止了外源蛋白的降解.例如,Wong等将拟南芥rbcS亚基的转运肽序列连接于杀虫蛋白基因之前,发现杀虫蛋白能够特异性地积累在转基因烟草的叶绿体内,外源蛋白总的积累量比对照提高了10～20倍.最近,叶梁、宋艳茹等也将rbcS亚基的转运肽序列连接于PHB合成相关基因之前,试图使基因表达产物在转基因油菜种子的质体中积累,从而提高外源蛋白含量.另外,Wandelt等和Schouten等将内质网定位序列（四肽KDEL的编码序列）与外源蛋白基因相连接,发现外源蛋白在转基因植物中的含量有了显著提高.显然,定位信号对于促进蛋白质积累有积极作用,但同一种定位信号是否适用于所有的蛋白还有待于进一步确定. 6 内含子在增强基因表达方面的应用 内含子增强基因表达的作用最初是由Callis等在转基因玉米中发现的,玉米乙醇脱氢酶基因（Adhl）的第一个内含子（intron 1）对外源基因表达有明显增强作用,该基因的其他内含子（例如intron8,intron9）也有一定的增强作用.后来,Vasil等也发现玉米的果糖合成酶基因的第一个内含子能使CAT表达水平提高10倍.水稻肌动蛋白基因的第三个内含子也能使报道基因的表达水平提高2～6倍.至今对内含子增强基因表达的机制不不清楚,但一般认为可能是内含子的存在增强了mRNA的加工效率和mRNA稳定性.Tanaka等人的多项研究表明,内含子对基因表达的增强作用主要发生在单子叶植物,在双子叶植物中不明显. 由于内含子对基因表达有增强作用,Mcelroy等在构建单子叶植物表达载体时,特意将水稻的肌动蛋白基因的第一个内含子保留在该基因启动子的下游.同样,Christensen等在构建载体时将玉米Ubiquitin基因的第一个内含子置于启动子下游,以增强外源基因在单子叶植物中的表达.然而,有研究指出,特定内含子对基因表达的促进作用取决于启动子强度、细胞类型、目的基因序列等多种因素,甚至有时会取决于内含子在载体上的位置.例如,玉米Adhl基因的内含子9置于Gus基因的5‘端,在CaMV35S启动子调控下,Gus基因的表达未见增强；当把内含子置于Gus基因3端,在同样的启动子控制下,Gus基因的表达水平却增加了大约3倍.由此可见,内含子对基因表达的作用机制可能是很复杂的,如何利用内含子构建高效植物表达载体,目前还缺乏一个固定的模式,值得进一步探讨. 7 多基因策略 迄今为止,多数的遗传转化研究都是将单一的外源基因转入受体植物.但有时由于单基因表达强度不够或作用机制单一,尚不能获得理想的转基因植物.如果把两个或两个以上的能起协同作用的基因同时转入植物,将会获得比单基因转化更为理想的结果.这一策略在培育抗病、抗虫等抗逆性转基因植物方面已得到应用.例如,根据抗虫基因的抗虫谱及作用机制的不同,可选择两个功能互补的基因进行载体构建,并通过一定方式将两个抗虫基因同时转入一个植物中去.王伟等将外源凝集素基因和蛋白酶抑制剂基因同时转入棉花,得到了含双价抗虫基因的转化植株.Barton等将Bt杀虫蛋白基因和蝎毒素基因同时转入烟草,其抗虫性和防止害虫产生抗性的能力大为提高（专利）.在抗病方面,本实验室蓝海燕等构建了包含β-1,3-葡聚糖酶基因及几丁质酶基因的双价植物表达载体,并将其导入油菜和棉花,结果表明,转基因植株均产生了明显的抗病性.最近,冯道荣、李宝健等将2～3个抗真菌病基因和hpt基因连在一个载体上,两个抗虫基因与bar基因连在另一个载体上,用基因枪将它们共同导入水稻植株中,结果表明,70%的R.代植株含有导入的全部外源基因（6～7个）,且导入的多个外源基因趋向于整合在基因组的一个或两个位点. 一般常规的转化,尚不能将大于25kb的外源DNA片段导入植物细胞.而一些功能相关的基因,比如植物中的数量性状基因、抗病基因等,大多成"基因簇"的形式存在.如果将某些大于100kb的大片段DNA,如植物染色体中自然存在的基因簇或并不相连锁的一系列外源基因导入植物基因组的同一位点,那么将有可能出现由多基因控制的优良性状或产生广谱的抗虫性、抗病性等,还可以赋予受体细胞一种全新的代谢途径,产生新的生物分子.不仅如此,大片段基因群或基因簇的同步插入还可以在一定程度上克服转基因带来的位置效应,减少基因沉默等不良现象的发生.最近,美国的Hamilton和中国的刘耀光分别开发出了新一代载体系统,即具有克隆大片段DNA和借助于农杆菌介导直接将其转化植物的BIBAC和TAC.这两种载体不仅可以加速基因的图位克隆,而且对于实现多基因控制的品种改良也会有潜在的应用价值.目前,关于BIBAC和TAC载体在多基因转化方面的应用研究还刚刚开始. 8 筛选标记基因的利用和删除 筛选标记基因是指在遗传转化中能够使转化细胞（或个体）从众多的非转化细胞中筛选出来的标记基因.它们通常可以使转基因细胞产生对某种选择剂具有抗性的产物,从而使转基因细胞在添加这种选择的培养基上正常生长,而非转基因细胞由于缺乏抗性则表现出对此选择剂的敏感性,不能生长、发育和分化.在构建载体时,筛选标记基因连接在目的基因一旁,两者各有自己的基因调控序列（如启动子、终止子等）.目前常用的筛选标记基因主要有两大类：抗生素抗性酶基因和除草剂抗性酶基因.前者可产生对某种抗生素的抗性,后者可产生对除草剂的抗性.使用最多的抗生素抗性酶基因包括NPTII基因（产生新霉素磷酸转移酶,抗卡那霉素）、HPT基因（产生潮霉素磷酸转移酶,抗潮霉素）和Gent基因（抗庆大霉素）等.常用的抗除草剂基因包括EPSP基因（产生5-烯醇式丙酮酸莽草酸-3-磷酸合酶,抗草甘磷）、GOX基因（产生草甘膦氧化酶、降解草甘膦）、bar基因（产生PPT乙酰转移酶,抗Bialaphos或glufosinate）等. 上面这些当中1、2、3、5、6都是值得注意的,特别是5,因为你要向细胞外分泌.骨架载体可以选择PB I121,然后你可以在上面改动基因型. 后面的就是克隆的步骤了,相对简单. 1 首先获得目的基因加酶切位点,连入改好的载体中. 2 将质粒转入大肠杆菌DH5a扩增 3 将扩增好的质粒转入植物细胞内进行表达 4 收集根细胞外培养基检测是否有该蛋白的表达和分泌. 至于改造载体那几个步骤要是答题的话简单说说就可以了,毕竟如果真的做出一个好载体都可以自己开公司了.

淮阴工学院为本科二批录取院校。淮阴工学院隶属于江苏省，是一所以工为主、理、管、文、法等多学科协调发展的本科院校。学校创建于1960年，原名淮阴师范学校，1984年更名为淮阴工学院。学校位于江苏省淮安市清河新区，校园占地面积2000余亩。学校现有工程学、理学、管理学、文学、经济学、法学、艺术学7个学科门类，设有66个本科专业，涵盖工、理、工程、管理、文、法、教育、艺术八大领域。学校拥有优美的校园环境、先进的教学设施和设备，为学生提供良好的学习和生活条件。师资力量：淮阴工学院拥有一支优秀的师资队伍，截至2021年，学校教职工总数约为1800人，其中教授、副教授占本科教师的60%以上，硕士生导师约占教师总数的38%。同时，学校引进了一大批海内外知名专家学者，包括两院院士、长江学者、国家杰出青年科学基金获得者等。学校注重师资队伍建设和教师培养，实施有针对性的“双带头人”培养计划、骨干教师培训、中青年教师培养等项目，不断提升教师的教学水平和科研能力。学校还鼓励教师积极参加各类学术会议和科研项目，加强国际学术交流，提高教师的国际视野和学术水平。

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