筛选转基因动物的新方法——PCR及其产物测序法

A New Method of Screening Transgenic Animal: PCR and Its Products Sequencing.

用特异性引物对肌球蛋白轻链 2启动子 (myosinlightchain 2 ,MLC2 ) 糜酶融合基因的转基因新生鼠鼠尾DNA进行PCR筛选 ,低熔点琼脂糖凝胶电泳回收阳性样品PCR所扩出的DNA条带 ,纯化后用同一对引物中的一个进行单引物PCR测序 ,与所转外源基因序列比较 ,进一步确定整合有外源基因的阳性鼠 .PCR及PCR产物测序法检测转基因动物具有操作方便 。

The primary detection of the MLC 2 chymase fusion gene transgenic mice was carried out by means of polymerase chain reaction(PCR) with two specific primers, which span across MLC2 promoter region and chymase structure gene region. The DNA electrophoretic bands of PCR products were recovered and purified , then sequenced by PCR sequencing method with one of the two primers. Final determination of the transgenic mice was conducted through comparing the sequencing results with sequences of transfered gene. This method is convenient, effective and specific.