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

作　　者：贾小霞[1] 张金文[1] 王汉宁[1] 吴文俊[1] 

机构地区：[1]甘肃农业大学农学院,甘肃兰州730070

出　　处：《兰州大学学报（自然科学版）》2008年第6期34-38,共5页Journal of Lanzhou University(Natural Sciences)

基　　金：科技部支撑项目(2007BAD52B08)资助.

摘　　要：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.

关 键 词：融合基因 几丁质酶基因 Β-1 3-葡聚糖酶基因 植物表达载体 

分 类 号：S572[农业科学—烟草工业] Q786[农业科学—作物学]