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Or you could ask the question, I have this restriction enzyme, at what regions on this plasmid will it cut?
你也可以这样问,我的这种限制性内切酶,会从质粒的哪个区域切开呢
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In this case, this particular restriction enzyme cuts symmetrically like this, but not at the same point.
在我们这种情况下,这个限制性内切酶总是这样对称地切割,但两条链切割的位点不一样
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This restriction enzyme was found in a natural source, it was found in a micro-organism called E.coli.
这个限制性内切酶源于自然界,是从名为大肠杆菌的微生物中发现的
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That restriction enzyme, there is a restriction enzyme that does that and it's MST-3.
确实存在这种限制酶,叫MST-3
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Most restriction enzyme also recognize symmetric sequences of DNA, GAATTC for example.
大多数限制性内切酶,也能识别DNA的对称序列,例如GAATTC
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If I cut both the plasmid and my DNA of interest with the same restriction enzyme I'm going to end up with the same sticky ends on both molecules.
如果用同一种限制性内切酶,来切割质粒和我感兴趣的DNA,在两个分子上就能得到同样的粘性末端
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If I took this same restriction enzyme and tried to cut the chromosomal DNA of a sickle patient, it wouldn't cut at that point because the wrong sequence is there.
而如果我用同样的限制酶,去切割病人的染色体DNA,那该酶是不会起作用的,因为基因序列不对
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Restriction enzymes are just a kind of enzyme, enzymes are protein molecules that make a chemical reaction go faster, and the chemical reaction that restriction enzymes do is cutting DNA.
限制性内切酶是一种酶,而酶是加速化学反应的蛋白质分子,限制性内切酶加速的化学反应,是切割DNA
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This particular restriction enzyme here recognizes this sequence,GAATTC.
这里的这种限制性内切酶,只识别序列GAATTC
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Well one way you could do it is by saying 'if I have this one base pair difference then this sequence is going to be cut by a specific restriction enzyme that recognizes the sequence CTGAGGA'.
一种方法是通过考虑,"如果我有这个不同的碱基对,那该序列必然会,被特定的限制酶识别并切割,这个特定的限制酶能识别,CTGAGGA "
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In a normal cell DNA that DNA in the normal cell - this DNA gets cut by the restriction enzyme, so the sickle gene ends up - so that hemoglobin gene ends up in two pieces.
一个正常细胞的DNA,正常细胞里的DNA,会被限制酶所剪切,因此镰状血红蛋白基因--,不对,是正常血红蛋白基因被切成两段
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Well the first step would be to cut open the plasmid with a particular restriction enzyme, and then what if I take that same restriction enzyme and I cut up the DNA that I'm interested in.
第一步是用某种限制性内切酶把质粒切开,然后用同一种限制性内切酶,切出我想要的DNA
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We're going to take a chromosomal DNA, we're going to digest it with this restriction enzyme, we're going to put it in this tube and run it on a gel, and we're going to see what results down here.
我们取一条染色体DNA,然后用限制酶进行分解,接着再放,在凝胶上进行电泳,之后看看结果如何
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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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