Chi-linker-Glu融合基因双价植物表达载体的构建及其相关鉴定

Construction of plant expression vectors containing Chi-linker-Glu fusion genes and their correlated identification

将木霉几丁质酶基因Chi和β-1,3-葡聚糖酶基因Glu通过8个中性氨基酸连接起来,形成Chi- linker-Glu融合基因.其中编码区基因长2376 bp,共编码792个氨基酸和一个终止子.将该融合基因克隆到中间载体pAHC25中,然后将整个ubi-chi-linker-glu-nos表达盒插入用ubi-bar-nos表达盒代替CaMV35S-gus-nos的高效植物表达载体pBI121中,构建了双价抗真菌基因的重组质粒pBIb-CG,并通过根癌农杆菌介导,采用叶盘法转化烟草.PCR扩增和PCR-Southern分析证明已将Chi-linker- Glu融合基因整合到烟草基因组中.

Chitinase genes and β-1, 3-Glucanase genes were connected by eight neutral amino acids, forming a fusion gene Chi-linker-Glu which had a coding region of 2 376 bp that encoded 792 amino acids and a terminator. The plant expression vector pBIb-CG was constructed by cloning the fusion gene to the intermedial vector pAHC25 and inserting ubi-chi-linker-glu-nos into pBI121 which replaced CaMV35S-gus- nos with ubi-bar-nos. By Agrobacteriura turner aciens-mediated genetic transformation, transgenic plantlets were obtained. It was proved that the fusion gene had been integrated into the tobacco genome by PCR amplification and PCR-Southern blotting analysis.