木瓜凝乳酶基因克隆及毕赤酵母分泌型重组表达载体构建

Gene cloning of chymopapain and construction of the secreting recombinantvector in Pichia pastoris

目的扩增木瓜凝乳酶基因 ,构建重组表达载体。方法提取木瓜组织RNA ,采用RT PCR方法扩增木瓜凝乳酶基因 ,测序并利用BLAST软件进行序列分析。利用基因重组方法构建重组表达载体。结果扩增得到木瓜凝乳酶基因。结论构建了可在毕赤酵母表达系统中表达天然木瓜凝乳酶蛋白的重组表达载体。

PurposeThe chymopapain gene was amplified, and the recombinant vector was constructed. MethodsRNA was separated from Carica papaya tissues. The chymopapain gene was amplified by RT-PCR. The gene fragment was sequenced,and the result was analyzed by BLAST program.ResultsThe interest gene was separated by RT-PCR. ConclusionThe recombinant vector was constructed.