摘要
为克隆和分析禽流感病毒(H9亚型)HN31株的NS基因的分子特征,通过RT-PCR方法扩增其NS基因,PCR产物与载体pMD19-TVector连接,转化大肠杆菌DH5α,经蓝白斑筛选,调取阳性菌落,摇菌过夜,提取质粒进行PCR鉴定和酶切鉴定,送阳性重组质粒测序。结果显示RT-PCR产物大小为919bp,HN31株的NS基因长度为890bp;其与H6、H9亚型的A型流感病毒的亲缘关系比H5、H7、H3、H1亚型流感病毒更近。
In order to study the molecular characteristics of the avian influenza virus （H9 subtypes） HN31 strain＇ s NS gene, its nucleotides were cloned into pMD19-T Vector by RT-PCR. The ligation product was transformed into the competent E.coli DHSct. The white colony was selected, and identified by PCR and EcoR I restriction enzyme digestion. The positive plasmid was sequenced. The results showed that the RT-PCR product was 919 bp, HN31 strain＇s NS gene length was 890 bp. Through the phylogenetic analysis, the kinship with H6, H9 subtype was closer than with HS, H7, H3, H1 subtype of influenza virus.
出处
《中国农学通报》
CSCD
2012年第2期53-56,共4页
Chinese Agricultural Science Bulletin
基金
国家"十一五"科技支撑计划"新城疫等禽用新型疫苗的研制与开发"(2006BAD06A08)
