发信人: royluo (roy), 信区: Biology
标 题: Re: 有人在细菌里表达过cysteine rich protein吗？
发信站: The unknown SPACE (Fri Jul 26 07:40:03 2002), 站内信件

Do you have a way to distinguish correctly folded protein? If you have,
just use HPLC to purify them from crude E.Coli expressed protein.
If you don't have one way to do that.
Refolding involves (1) denaturing and reducing (2) suddently dilute
the sample extensively to mild condition (3) get the soluble protein.
Try to search pubmed for methods or refer to any classic protein biochemistry
protocol book. Note: the yield is low(less than 10% of the crude protein
will be correctly refolded).

Good luck!

【 在 scoop (大铲) 的大作中提到: 】
: 我的蛋白不大，看来还是可以用简单方法解决的了。那么refolding又有哪些方法呢？
: 【 在 royluo (roy) 的大作中提到: 】
: : If your protein is big or an enzyme, try 5.
: : If your protein is not big , try to do refolding after expression in E.Coli.
: : Usually, you need to have a way to purify the correctly folded protein
: : with correctly disulfide bonds. 123 only make things complicated, and don't
: : really help too much.
: 的困难在于细菌的cytoplasm 是还原性，disulfide bonds难以形成。所以有几个解决方
: 案：


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