单纯疱疹病毒1型解螺旋酶-引物酶编码基因重组杆状病毒载体的构建与鉴定

Construction and identification of recombinant baculovirus vectors of genes encoding helicase-primase of HSV-1

目的:分别构建含单纯疱疹病毒1型(HSV-1)解螺旋酶-引物酶编码基因UL5、UL52与UL8的重组杆状病毒载体。方法:PCR扩增UL5、UL52与UL8基因全长序列,分别克隆至转移载体pFastBac1,转化E.coliDH10Bac,通过转座作用制备分别带有3种目的基因的重组杆状病毒穿梭载体。结果:克隆的各目的基因序列与GenBank中收录序列一致,各目的基因正确重组至杆状病毒穿梭载体Bacmid。结论:成功构建分别插入HSV-1 UL5、UL52与UL8基因的重组杆状病毒载体,为在昆虫细胞内表达组装HSV-1解螺旋酶-引物酶以及建立靶向该酶复合体的药物筛选平台奠定基础。

Objective：To construct the recombinant baculovirus vectors of UL5, UL52 and UL8 respectively, genes encoding the helicase - primase of herpes simplex virus type 1 （ HSV - 1 ）. Methods： Desired genes were obtained by PCR reaction and were inserted into pFastBac! vectors, respectively. Recombinant pFastBacl vectors of each gene were transformed into E. coli DH10Bae to generate recombinant baculovirus vectors through transposition. Results： Sequence analysis revealed that the cloned gene segments were consistent with that of UL5, UL52 and UL8 of HSV - 1 in GenBank. Each gene was correctly recombined into baculovirus vector Bacmid. Condusion：The recombinant baculovirus vectors separately containing UL5, U L52 and U L8 were constructed successfully, laying foundation for the expression and assembly of helicase - primase in insect cells and for the establishment of anti - HSV - 1 drug screening platform targeting this enzyme complex.