鸡传染性喉气管炎病毒长葛株gE全基因的克隆与序列分析

Cloning and Sequence Analysis of gE Gene of Infectious Laryngotracheitis Virus Isolated from HN Changge Strain

[目的]防治鸡传染性喉气管炎,减少该病对养鸡业造成的损失。[方法]参考GenBank收录的传染性喉气管炎病毒gE基因序列设计并合成1对引物,以ILTV河南长葛株DNA为模板,PCR扩增出一条1 550 bp的特异性条带,并克隆至pGEM-T载体上。经PCR扩增、酶切鉴定,得到含gE基因重组质粒,并对重组阳性质粒进行序列测定。[结果]与GenBank收录的传染性喉气管炎病毒gE基因和其他部分疱疹病毒gE基因序列进行比较,发现传染性喉气管炎病毒间的核苷酸和氨基酸的同源性分别为99.9%、99.8%;与其他动物疱疹病(MDV、CHV、PRV、EHV的gE基因)毒核苷酸的同源性分别为25.6%、23.9%、28.3%、26.1%。[结论]不同ILTV毒株之间gE基因差异甚小,gE基因比较保守,但与其他动物疱疹病(MDV、CHV、PRV、EHV的gE基因)毒核苷酸的同源性较低。该研究为gE基因的功能研究提供了依据。

According to published ILTV gE gene sequences,one pair of specific primers was designed and synthesized.The genomic DNA of ILTV HN Changge strain,which was isolated from Henan Changge,was used as template.Full-length gE gene was amplified by polymerase chain reaction（PCR） and cloned into the pGEM-T easy vector.The gene was sequenced and the open reading frame of 1 550 bp was determined.Compared ILTV gE gene sequence of HN Changge strain with the corresponding sequence of other strains of ILTV and other partial herpesvirus.The homology of the nucleotide sequence was 99.9% and the homology of the amino acid was 99.8% between ILTV strains,but the homology of the nucleotide sequence was only 25.6%,23.9%,28.3% and 26.1% with other animal herpesvirus strains.Comparative sequence analysis showed the HN Changge strain had a high homology with other strains of ILTV.It indicated that ILTV gE gene was conserved gene.