猪轮状病毒地方株JL94株VP4基因的克隆与序列分析

Cloning of VP4 gene from porcine rotavirus JL94 isolate and homologous comparison with reference strains

根据GenBank已发表的猪轮状病毒VP4基因保守序列,利用Oligo4.1软件设计—对引物扩增猪轮状病毒地方株JL94株VP4全基因序列;并将扩增产物与pMD-18T载体连接,经鉴定后将重组质粒进行测序并与国外分离株CRW-8株、BEN-307株进行核苷酸和氨基酸序列同源性比较分析。结果表明:JL94株和CRW-8株、BEN-307株VP4全基因片段核苷酸同源性和氨基酸同源性分别为96.98％和98.05％,说明JL94株与CRW-8株、BEN-307株属于同一VP4血清型。本研究为进一步研制诊断性抗原和病毒重组活载体疫苗奠定了基础。

A pair of primers according to the published cDNA sequence of porcine rotavirus VP4 gene were designed, which were used in cloning and sequence analysis of VP4 gene from porcine rotavirus JL94 isolated in China. The full length gene encoding outer capid protein VP4 was amplified from the native JL94 isolate of porcine rotavirus by reverse transcription polymerase chain reaction(RTPCR). The cDNA of VP4 gene was inserted into pMD-18T vector and identified by sequencing. The result of its homology comparison with CRW-8 strain and BEN-307 strain is 96.98 % and 98.05 % respectively.