A型流感病毒株核蛋白基因的克隆与重组质粒的构建  

Cloning of Influenza A Nucleoprotein and Constructing the Recombinant Plasmid

作　　者：马一君[1] 藏文巧[2] 王红炜[1] 胡军[2] 杜献堂[2] 

机构地区：[1]河南中医学院基础医学院病原与免疫学科,郑州450008 [2]郑州大学基础医学院微生物与免疫学教研室,郑州450052

出　　处：《热带医学杂志》2006年第3期287-289,共3页Journal of Tropical Medicine

摘　　要：目的克隆A型流感病毒株Swine/Henan/703/2001(H3N2)的核蛋白(NP)基因序列并构建重组质粒。方法采用逆转录聚合酶链式反应技术从病毒株基因组中扩增出编码核蛋白的基因序列,克隆至pGEM-TEasy载体,经蓝白筛选、菌落PCR及酶切鉴定挑取阳性克隆并测序。结果利用RT-PCR扩增到目的基因片段,并将其克隆至T载体。结论成功构建重组质粒pGEM-TEasy-NP,为NP相关功能及流感疫苗的进一步研究奠定实验基础。Objective To clone the gene fragment encoding nueleopmtein （NP）of influenza h/Swine/Henan/ 703/2001 （H3N2）and construct the recombinant plasmid. Methods The sequence encoding NP was amplified by reverse transcriptase-polymerase chain reaction from the virus RNA genome. The gene was then cloned into pGEM-T Easy vector. Positive clones were identified by blue-white screening and colony PCR, and the gene was obtained by digesting the plasmid with restriction endonuclease. The sequence of NP was confirmed by DNA sequencing. Results The target gene fragment was amplified by RT-PCR and inserted into T vector. Conclusion Recombinant plasmid pGEM-T Easy-NP was constructed suceessfuUy. It will be used in the functional studies of NP and in the development of influenza virus subunit vaccine.

关 键 词：A型流感病毒 核蛋白 克隆 重组质粒 

分 类 号：R511.7[医药卫生—内科学]