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If you have DNA fragments, so this is DNA that you've cut up into fragments using restriction enzymes for example.
如果你有DNA片段,例如有一段你用限制酶,切割成片段的DNA
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Not every plasmid in your test tube is going to have the right gene inserted in the right way.
不是试管里的每个质粒,都会与正确的DNA片段重组
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You have a fragment of some kind of DNA that you would like to clone or make many copies of.
你有了某种DNA的片段,想要克隆,复制它
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They're not whole genomes, but they're some small piece of DNA maybe the size of a protein.
不是整个基因组,而是小的DNA片段,但也许与蛋白质分子一样大
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Every cell in your body is only using a fraction of the DNA that's available to it.
每个细胞只用到了,决定该细胞生物功能的DNA片段
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How did we put our gene fragments into this plasmid DNA in order to make multiple copies of it, or to clone the gene?
如何把基因片段整合到质粒DNA上,来获得大量拷贝,或者说来克隆基因呢
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So bacteria that get an unloaded plasmid are going to have resistance to antibiotics.
得到无DNA片段质粒的细菌,就会对抗生素有抗性
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That's a really good question because if I open this up, why wouldn't it just reform with itself, why would it want to have this in here?
这是个很好的问题,如果我把质粒打开,为什么它不会自己合上,为什么它会与DNA片段重组呢
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If I cut up just DNA from the chromosome, what's called genomic DNA, then I'm going to have both exons and introns within the fragments that I create.
如果我切割的恰好是染色体上的,基因组DNA,那我切成的片段上,就会同时有外显子和内含子
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If you start with only one, you have two pieces of DNA, then you'll get 2 to the Nth fragments after N cycles because each cycle you're doubling the number.
如果你从仅仅一个DNA开始,你有两条DNA链,经过N次循环后,就得到二的N次方个DNA片段,因为每次循环都使其数量翻倍
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So the DNA is going to get spread out on this gel according to size, with the small ones going farther and the large ones not going as far.
因此DNA会根据大小,在凝胶上分散开,小的片段会快一些,大的片段不那么快
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Now if I can stain in some way, if I can somehow label the DNA fragments that I'm interested in, I could find out where those fragments are on this gel.
那如果我有某种染色的方法,对我感兴趣的DNA片段进行标记,我就能知道这些片段在凝胶板上的位置
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Some are going to reform and some are going to reform with the gene in, and some probably aren't going to reseal at all under the conditions that I've used.
有些质粒会自己合上,有些质粒会与DNA片段重组并合上,还有的在我提供的条件下,也根本合不上
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One of the forms of processing that happens, that's very important in human gene expression is that some of the sections of the DNA molecule are not really necessary for describing the protein.
在人类的基因表达中,有一种非常重要的加工形式,那就是在DNA分子中有一些片段,对于编码蛋白质而言毫无用处
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If you cut and you open up a segment of DNA then you've left these sticky ends, for example and these sticky ends are capable of recognizing each other by the process of hybridization.
如果你进行切割打开一个DNA片段,就会留下,比如说,这些粘性末端,这些粘性末端能在杂交的过程中互相识别
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Let's assume that we have this plasmid cloning vector and we have some pieces of DNA that we would like to put into a plasmid that we would like to make copies of.
假设我们已经有了克隆质粒载体,也有了我们想要将之,整合到质粒上的某些DNA片段,同时也是我们所要复制的DNA片段
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The first step is to take the DNA fragments that we're interested in and put them into this vector by basically cutting open the double stranded DNA and inserting the gene that we like in the region where we've cut.
第一步是把我们感兴趣的DNA片段,整合到这个载体里,基本的方法是切开质粒这个双链DNA,把我们想要的基因插入切开的区域
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Now if I put them in contact with one another, the plasmid that's been opened and fragments of the DNA - special fragments that I've produced with the same restriction enzyme, they'll have the same sticky ends, they will naturally hybridize with one another.
如果我让它们互相接触,已经切开的质粒和DNA片段--,这DNA片段是我已用同种限制性内切酶,处理过的特别片段,这两者会有相同的粘性末端,它们会自然地相互杂交
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I'm sure some of you have dealt with electrophoresis gels in the past and it's really just a slab of something like Jell-O where you can put a sample at one end of the Jell-O, your sample of fragmented DNA.
我相信在座肯定有些已经接触过电泳凝胶,真的挺像一块板状果冻的,你可以把一段样本放在"果冻"的一端,你的DNA片段样本
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The answer is it will want to reform with itself, and if I have these in solution then how many reform with itself and how many reform with the molecule I'm interested probably depends on the relative concentrations of both in the solution and what conditions I have it at.
答案是质粒有可能会自己闭合,如果质粒和我感兴趣的DNA片段,都在溶液里那有多少质粒会自己合上,有多少与我感兴趣的DNA重组呢,这大概取决于两者在溶液里的相对浓度,和我让溶液处在什么条件下
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