木瓜凝乳酶基因的克隆及序列分析

Cloning and Sequence Analysis of Chymopapain Gene

通过设计一对特异性的引物,采用RT-PCR方法从未成熟的木瓜组织中扩增得到木瓜凝乳酶基因,并将其重组到pPIC9K载体中,转化大肠杆菌并筛选阳性克隆,序列测定并利用BLAST软件进行核酸及氨基酸序列相似性分析,结果表明:通过序列组成及特征结构分析,扩增得到的基因为木瓜凝乳酶基因。

The chymopapain gene was amplified using special primers from Carica Papaya tissues by RT-PCR and wa s inserted into plasmid pPIC9K. The recombined plasmid was transformed into E. coli TOP10, and positive transformant was screened. The interest fragment was sequenced, and the nucleic acid sequence and amino-acid sequence encoded by the gene were compared using BLAST program online. The result show that the amplifi ed cDNA fragment is a novel chymopapain gene based on the sequence constitute an d analysis of character construction. [WT5HZ]