分子杂交法鉴定人睾丸cDNA重组片段

IDENTIFICATION OF HUMAN TESTIS cDNA RECOMBINATION BY MOLECULAR HYBRIDIZATION

鉴定cDNA表达文库构建是否成功的一个重要指标是cDNA片段与噬菌体载体的重组率,为能精确地反映出我们所构建的人睾丸cDNA表达文库的重组率,我们在运用遗传显色法检测的同时,进一步运用分子杂交法进行了测定.结果显示,运用遗传显色法测定的重组率为66%,分子杂交法测定的重组率为51%.表明所构建文库具有较高的重组率,并提示遗传显色法测出的重组率中有部分是因为基因休克导致的假阳性.运用分子杂交法虽然方法较复杂,但准确性高,应该提倡使用.

Recombination rate is one of the important remark for the identification of cDNA Library. In our laboratory, genetic method and molecular hybridization were used for the determination of recombination rate. The results showed that the recombination rate of the cD-NA library was rather high and indicated that the recombination rate was 66% determine by genetic method and 51% by molecular hybridization. The higher rate determined by genetic method may partly be caused by gene shock which can cause false-positive. Although the molecular hybridization is complex in method, it is accurate and suited for wide use.