鸡传染性喉气管炎病毒王岗株gC和gD基因的克隆及序列分析

Cloning and Sequencing of Genes encoding the Glycoprotein C(gC and D(gD) of Infectious Laryngotracheitis Virus(ILTV) Wanggang Strain

根据已发表的传染性喉气管炎病毒 ( ILTV) SA-2株的核酸序列 ,设计了 1对引物 ,以 ILTV中国王岗株 DNA为模板 ,PCR法扩增出 1条 1 .3 9kb的基因片段 ,将扩增产物插入到真核表达质粒 p CRTM 3 -Uni,得到重组质粒 p TA-g D。另将克隆到 p Bluescript SK质粒中的 g C基因酶切后 ,插入到 p CRTM 3 -Uni载体 ,得到重组质粒 p TA-g C。经电泳分析、酶切、PCR鉴定后 ,进行序列测定。序列分析表明 ,ILTV王岗株 g C基因的编码序列与 SA-2株的核苷酸同一性为 99%,而王岗株 g D基因的编码序列在其 C端多了 1 7个核苷酸 ,其余部分的同一性为 97%。鉴于 g C、g D基因在诱导机体产生对 ILTV的免疫应答中的作用 ,g C、g D基因的克隆及其真核表达质粒的构建 ,为

A fragment containing gD gene of ILTV Wanggang strain was amplified by polymerase chain reaction and cloned into pCR TM 3 Uni vector and sequenced. A recombinant plasmid containing the coding sequence of gC gene ILTV of Wangggang strain was cleaved with restriction endonuclease Eco RI, and a 1 292 bp fragment of gC gene was ligated into the Eco RI site of pCR TM 3 Uni vector and sequenced.Sequence analysis indicated that gC gene of ILTV Wanggang strain displayed 99% nucleotide identities with SA 2 strain, and the gD gene of ILTV Wanggang strain was 17 bp longer than that of SA 2 strain in the C terminus,and the rest part of the gene from the two strains was almost the same.