杆状病毒ac30基因的序列分析及其功能
Sequence analysis and functional study on baculovirus ac30 gene

通过生物信息学初步分析苜蓿丫纹夜蛾核多角体病毒（Autographa californica multiple nucleopolyhedrovirus,AcMNPV）基因组第30开放阅读框（ac30）及其同源基因的特征，并在此基础上利用ET-重组技术构建ac30 缺失突变体,探讨其在AcMNPV感染Sf-9细胞时的功能。结果表明：ac30的开放阅读框(ORF)全长为1392 bp，GC 含量为43%，编码氨基酸463个，预计分子质量为54.7 ku。从转染或感染的病毒生长曲线变化趋势来看, ac30的缺失对病毒产生有感染力的BV粒子没有影响，说明ac30基因是AcMNPV病毒在Sf-9细胞复制中的非必需基因。
Autographa californica multiple nucleopolyhedrovirus(AcMNPV) ac30 (orf30) is a highly conserved gene in lepidopteran baculovirus whose function is unknown. ac30 is located between nt 24315 to nt 25706 in AcMNPV genome, and encodes a putative protein of 463 amino acids with a predicted molecular mass of 54.7 ku. To determine the role of ac30 in the baculovirus life cycle, an ac30 knockout virus containing the AcMNPV genome was generated by using AcMNPV bacmid bMON14272 via ET-recombination in Escherichia coli. To rescue the phenotype of the ac30 knockout, an ac30-repaired virus was constructed in which an ac30 gene copy with its native promoter and terminator element was inserted into the polyhedron (polh) locus of the ac30 knockout bacmid by transposition.Sf-9 cells were transfected withwt bacmid,ac30 knockout bacmid or the repair bacmid,respectively.All of the three virus bacmids could generate infectious virus. After infection of Sf-9 cells with wt virus, ac30 knockout virus or ac30 repair virus, all of the three viruses showed similar infection pattern.All these results indicated that ac30 gene was not essential for virus propagation in AcMNPV life cycle.