人白介素-12植物表达载体的构建  

Construction of plant expression vector of human interleukin-12

作　　者：邵敏[1] 周鹤峰[2] 葛正龙[3] 

机构地区：[1]遵义医学院珠海校区生物化学教研室,广东珠海519041 [2]遵义医学院珠海校区生物工程教研室,广东珠海519041 [3]遵义医学院生物化学与分子生物学教研室,贵州遵义563003

出　　处：《第四军医大学学报》2007年第10期868-870,共3页Journal of the Fourth Military Medical University

基　　金：贵州省科委重大攻关项目(2004NGZ002)

摘　　要：目的构建人白介素-12(hIL-12)基因的植物表达载体.方法采用PCR技术从质粒pCA13-hIL-12中扩增hIL-12基因,SmaI,SacI分别酶切hIL-12基因和植物表达载体pBI121,回收,T4DNA连接酶连接得到重组质粒pBI121-hIL-12,双酶切和DNA测序鉴定后,通过三亲杂交法将表达载体pBI121-hIL-12分别导入根癌农杆菌EHA105和GV3101,用PCR法进行鉴定.结果双酶切及DNA序列测定证实hIL-12已插入到表达载体pBI121中,PCR扩增表明构建的重组表达载体pBI121-hIL-12成功转入根癌农杆菌EHA105和GV3101中.结论成功构建了植物表达载体pBI121-hIL-12,为进一步利用转基因植物生产hIL-12奠定了实验基础.AIM： To construct the plant expression vector of human interleukin-12 （ hIL-12 ） gene. METHODS ： Polymerase chain reaction （PCR） was used to amplify hIL-12 gene from the plasmid pCA13-hIL-12. Both hIL-12 gene and the plant expression vector pBI121 were digested by SmaI and SacI respectively, recovered, and ligated through T4DNA ligase to obtain pBI121- hIL-12. The recombinant plasmid was checked by restriction enzyme digestion and DNA sequencing, and then it was transferred into Agrobacterium Turnefaciens EHA105 and GV3101 by triparental mating. RESULTS： Restriction enzyme digestion and DNA sequencing indicated that hIL-12 gene was inserted into the expression vector pBI121, and PCR proved that plant expression vector pBI121-hIL-12 was successfully transferred into Agrobacterium Turnefaciens EHA105 and GV3101. CONCLUSION ： Successful construction of the plant expression vector pBI121-hIL-12 lays an experimental foundation for using the transgenic plants to produce hIL-12.

关 键 词：人白细胞介素-12 植物表达载体 载体构建 根瘤菌属 

分 类 号：Q78[生物学—分子生物学]