人表皮生长因子基因重组腺病毒穿梭质粒的构建和鉴定  

Construction and identification of a recombinant adenoviral shuttle plasmid expressing hEGF gene

作　　者：王巧稚 韩艺 邹礼乐 赵宏贤 梅欣明 

机构地区：[1]四川医科大学组织学与胚胎学教研室,四川泸州646000

出　　处：《泸州医学院学报》2016年第1期43-45,共3页Journal of Luzhou Medical College

基　　金：四川省卫生厅科研项目(NO:90192)

摘　　要：目的:构建含人表皮生长因子(human epidermal growth factor,hEGF)基因的重组腺病毒穿梭质粒,为制备表达EGF基因的重组腺病毒奠定基础。方法:查找Gen Bank中hEGF的基因序列,通过化学方法进行全基因合成得到信号肽+hEGF基因片段,将sp-hEGF基因装载到pUC57质粒,双酶切后与经过同样处理的腺病毒穿梭质粒pYr-ads-6相连,即构建成p Yr-ads-6-hEGF质粒,行双酶切及测序鉴定。结果 :sp-hEGF成功插入腺病毒穿梭质粒pYr-ads-6中,测序鉴定与Gen Bank中的序列一致。结论:hEGF重组穿梭质粒构建成功。Objective： To construct a recombinant adenoviral shuttle plasmid expressing hEGF gene to deliver hEGF gene effectively. Methods： sp-hEGF cDNA based on the Gen Bank was chemically synthesized and inserted into the plasmid puc57. The plasmid was digested with Nde I and Hind III, and the double-digested fragment was then cloned into adenoviral shuttle plasmid pYr-ads-6, forming pYr-ads-6-sp-hEGF. The recombinant adenoviral plasmid was verified by double digestion and DNA sequencing. Results： sp-hEGF was successfully synthesized and cloned into adenoviral shuttle plasmid pYr-ads-6. Conclusion： The recombinant adenoviral plasmid carrying hEGF gene is successfully constructed.

关 键 词：HEGF 腺病毒 穿梭质粒 信号肽 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学] R-33[医药卫生—基础医学]