人干扰素α-2b基因在烟草中的表达研究  

Expression of Human Interferon α-2b Gene in Tobacco(Nicotiana tabacum L.)

作　　者：崔翠菊[1] 赵稳[1] 周璇[1] 谭翼[1] 李肖[1] 杨广笑[1] 汪越胜[1] 何光源[1] 

机构地区：[1]华中科技大学生命科学与技术学院中英HUST-RRes基因工程和基因组学联合实验室,科技部国际科技合作基地(基因工程),教育部分子生物物理重点实验室,武汉430074

出　　处：《植物科学学报》2011年第1期81-86,共6页Plant Science Journal

基　　金：国家科技重大专项--优质转基因小麦新品种培育(2008zx08002-004);973预研项目(2008CB117014)

摘　　要：应用PCR的技术从质粒pAIFN中扩增人干扰素α-2b(Human interferon α-2b,HuIFN α-2b)编码基因,将其连接到pBI121双元载体构建植物真核表达载体pBIFN;用冻融法将该载体转染根癌农杆菌LBA4404;并用叶盘浸染法转化烟草叶片,经转化的烟草叶片的组织培养,诱导愈伤获得再生植株。通过应用PCR,RT-PCR,Wes-tern blot和WISH/VSV方法检测获得的烟草再生植株,结果表明HuIFN α-2b基因已成功整合进烟草核基因组并表达出具有活性的HuIFN α-2b蛋白。本文对HuIFN α-2b基因在烟草核系统中的表达进行了研究,为进一步在烟草叶绿体系统中该基因的表达研究奠定了基础。Polymerase Chain Reaction（PCR）amplifies the human interferon α-2b（HuIFN α-2b） gene encoding region from the plasmid pAIFN.The encoding region sequence was liga-sed into the pBI121 vector to construct plant eukaryotic expression vector pBIFN.The freeze-thaw method was used to transfect the vector pBIFN into Agrobacterium tumefaciens LBA4404 and then the leaf disc dip method was used to transform the A.tumefaciens LBA4404 embo-died with pBIFN into the tobacco（Nicotiana tabacum L.）leaf explants and plants were regene-rated from cultured transformed tobacco-leaf explants.The molecular analysis methods of PCR,RT-PCR,Western blot and the WISH/VSV were applied to confirm the regenerated tobacco plants and the results showed that the HuIFN α-2b gene was successfully integrated into the tobacco nuclear genome and HuIFN α-2b protein was expressed with activity.This paper documents the expression of the HuIFN α-2b protein in tobacco nuclear transformation system and implies that the potential application of HuIFN α-2b gene in tobacco chloroplast transformation system is feasible.

关 键 词：烟草转基因 人干扰素Α-2B 根癌农杆菌 

分 类 号：Q26[生物学—细胞生物学] Q943.1