利用三引物PCR和同尾酶连接的策略实现多基因片段的重组

Multi-gene fragments recombination by TP-PCR and isocaudamer ligation

在构建质外体CaM结合小肽中间载体的过程中,采用了三引物PCR和同尾酶连接法进 行DNA重组,构建了SLR1 BPs信号肽 CaN Cmyc的融合基因的一连体和CaN二连体。结果表 明,三引物PCR方法对多个小基因片段的重组操作方便快速,比酶切、连接方法更具优越性。 而同尾酶连接方法是连接多拷贝基因的有效方法。

In the process of constructing an intermediate vector, the Tri-Primer-PCR method was used in the construction of a one CaN vector and the isocaudamer enzyme ligation method was used in constructing a two CaN vector. The results showed PCR ligation was superior to enzyme digestionand ligation, and isocaudamer enzyme ligation was a practical way for the ligation of multi-copy foreign genes.