发信人: Ras (很颓的猪。), 信区: Biology
标 题: Re: Yeast two hybrid system?
发信站: The unknown SPACE (Wed Sep 13 14:01:36 2000), 转信



example:

--------------------- ----------------------------
| bait | activator| + |fragment A | reporter gene |
--------------------- ----------------------------

Bait ait is the gene u r intestered in, and is attached to a
transcription activator. fragment A is one of those from a DNA library
which is attached to a reporter gene.

Basically you transfect both the bait construct and library construct
into yeast (a and alpha respectively) and let the mate (each yest should have
only one of those library DNA fragment). If the bait can bind with one of
the fragments, then the transcription activator can be brought to the reporter
gene and transcribe the reporter gene. The expression of the reporter gene
will lead to blue color of the specific yeast colony. Then u pick the blue
colony, grow up culture, and purify DNA from it. You got the DNA that can
interact with your bait.


【 在 roche (晚安！) 的大作中提到: 】
: 谁能给我讲讲一个具体的例子。
: 我知道是用来检测两个蛋白质是否bind的， 但想知道具体的经典实例。
: 时间不多了，望刚复习过分子生物的朋友帮忙，(当初我也都知道，
: 现在忘了个干净，到哪儿找都不知道了 ：(((


--
一年老一年，一日没一日，
一秋又一秋，一辈催一辈，
一聚一离别，一喜一伤悲。
一榻一身卧，一生一梦里。
寻一夥相识，他一会咱一会，都一般相知，
吹一回，唱一回。

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