人白介素-12植物表达载体的构建

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.

Construction of plant expression vector of human interleukin-12

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 Tumefaciens EHA105 and GV3101 by tri-parental 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 Tumefaciens 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.