摘要
目的克隆甲型流感病毒H1N1亚型血凝素HA基因,并对其进行序列分析。方法利用TRIzol Rea-gent法提取甲型流感病毒H1N1亚型的总RNA,RT-PCR后以其cDNA为模板PCR扩增HA基因,经酶切、连接后,构建pET 28a(+)-HA重组质粒。利用生物信息学工具对HA的核酸序列和蛋白功能结构进行分析。结果成功构建了pET 28a(+)-HA重组质粒,经序列比对,与GenBank上公布的HA基因序列同源性达到99%。结论对HA基因进行克隆及生物信息学的分析,为深入研究HA在病毒侵入细胞及抗原变异中的作用机制及后续HA抑制剂的体外筛选奠定基础。
Objective To clone influenza A virus subtype H1 N1 hemagglutinin (HA) gene, and do sequence analy- sis. Methods The total RNA was isolated by TRIzol reagent from influenza A virus subtype H1N1, the cDNA was gained by RT-PCR and then used as a template for PCR, amplified HA gene. Recombinant plasmid pET 28a( + )-HA was con- structed. To analyze the nucleic acid sequence and properties of HA by bioinformatics analysis software. Results The re- combinant plasmid pET 28a( + )-HA was constructed, sequence alignment result showed that the sequence shares 99% of sequence homology with the HA reported in GenBank. Conclusion The cloning and bioinformatics analysis of HA lay a foundation for the deeply study of its role in virus replication cycle and the in vitro screening of HA inhibitors.
出处
《山东医药》
CAS
2013年第4期1-3,7,共4页
Shandong Medical Journal
基金
国家自然科学基金广东联合基金(U0632010)
广东高校皮肤黏膜释药制剂产学研结合示范(暨研究生创新培养)基地(cgzhzd0905)
关键词
流感病毒
H1N1亚型
血凝素
克隆
序列分析
influenza virus
subtype H1N1
hemagglutinin
cloning
bioinformatics analysis