原核注射产生转基因小鼠中的缺失整合

The Deletion Integration of Transgenic Mice Produced by Pronucleus Injection

用心脏特异性启动子肌球蛋白轻链－２（ｍｙｏｓｉｏｎｌｉｇｈｔｃｈａｉｎ－２）ＭＬＣ２和糜酶结构基因相拼接构建ＭＬＣ２－糜酶融合基因，通过原核注射法产生转基因小鼠。用特异性引物对新生鼠鼠尾ＤＮＡ进行ＰＣＲ扩增初选，Ｓｏｕｔｈｅｒｎ印迹杂交确定整合有外源基因的阳性鼠。低熔点琼脂糖凝胶电泳回收阳性鼠ＰＣＲ扩增的ＤＮＡ片段，经测序后，与所转外源基因序列比较，发现缺失整合现象。

The MLC 2 chymase fusion gene was constructed through splicing MLC 2 promoter sequence and cloned chymase structure gene.Transgenic mice carrying MLC 2 chymase fusion gene were generated by means of pronucleus injection.Screening for transgenic mice was carried out by PCR amplification with a pair of specific primers,Southern blot hybridization and sequencing of the PCR products. Deletion integration of transgenic mouse was found when compared the sequencing result with the transgenes.This study provids a new insight into behaviour of foreign DNA fragments integrating into host chromosome.