摘要
目的:通过构建人干扰素α-2b(hIFNα-2b)基因的植物表达载体,为后期将其导入胡萝卜愈伤组织中作准备。方法:采用PCR技术从人基因组DNA扩增hIFNα-2b编码基因及全长基因,将其克隆于pMD19-T载体中,双酶切hIFNα-2b基因及植物表达载体pBI121,回收目的片段,T4DNA连接酶连接得到植物表达载体pBI121-hIFN。采用三亲交配法将后者导入根瘤农杆菌。结果:经PCR检测,双酶切及DNA序列测定表明hIFNα-2b编码基因及全长基因已分别插入植物表达载体pBI121中,重组表达载体pBI121-IFN已成功转化根瘤农杆菌LBA 4404。结论:成功构建了植物表达载体pBI121-hIFN并转入根瘤农杆菌。
Objective：To construct plant expression vector containing human interferon alpha- 2b （hIFNα -2b） gene, which was to trans- form into carrot callus. Method：The human interferon alpha -2b coding sequence and full -length sequence were amplified from human genomic DNA by PCR, cloned into vector pMD19 -T, hIFN -α-2b genes and plant expression vector pBI121 were degested into pieces, then to recover the target fragments and to construct the new plant expression vector pBI121 -IFN by T4 DNA Ligase, which was trans- formed into Agrobacterium tumefaciens LBA 4404 by Triparental mating method. Result：According to the results of PCR, restriction enzyme digestion and DNA sequencing,it showed that the hIFNα- 2b genes （coding sequence and full -length sequence） were inserted into the plant expression vector pBI121 ,The recombinant expression vector pBI121 - IFN had been successfully transformed into Agrobacterium tu- mefaciens LBA 4404. Conclusion ：The plant expression vector pBI121 -IFN was successfully constructed,which brought the possibility for the further use of producing human interferon by transgenic plant.
出处
《生物技术》
CAS
CSCD
北大核心
2014年第1期8-12,共5页
Biotechnology
基金
贵州省科学技术基金项目(黔科合J字[2010]2151号)资助
关键词
人干扰素Α-2B
植物双元表达载体
转基因
根瘤农杆菌
Human interferon alpha- 2b
Plant binary expression vector
Transgene
Agrobacterium tumefaciens
