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传染性喉气管炎病毒烟台株gE基因的克隆与序列分析
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摘要:
根据传染性喉气管炎病毒(ILTV)gE基因序列设计并合成1对引物,以烟台株为模板,用PCR法扩增出1条1.5 kb的基因片段,并将其克隆至pMD18-T载体中。经电泳分析、酶切鉴定,将得到的重组质粒命名为pMD-gE并进行序列测定。将测序结果与ILTV美国标准攻毒毒株和中国王岗株,BHV-1,EHV-1,FHV-1,HHV-2,HHV-3,HVT,MDV-1,MDV-2,PRV的gE基因比较,发现核苷酸和氨基酸的同源性分别为99.7%~11.9%和99.4%~15%。结果表明,不同ILTV毒株之间gE基因还是比较保守的,但不同疱疹病毒之间,gE基因的同源性较低。
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Cloning and Sequence Analysis of the Glycoprotein E (gE) Gene of Infectious Laryngotractheitis Virus Yantai Strain
Abstract:
A pair of primers flanking the gE gene were designed according to the published nucleotide sequence of the ILTV.The Glycoprotein E (gE) gene of the wild Chinese ILTV Yantai strain was obtained by polymerase chain reaction and was cloned into pMD18-T vector.Based on the gel electropheresis and digestion with restrict enzyme,the identified positive recombinant plasmid was named pMD-gE and was sequenced.The results of sequence analysis showed that the homology of the gE gene and the deduced amino acid of ILTV Yantai strain with that of ILTV USA challenge strain,ILTV China Wanggang strain,BHV-1,EHV-1,FHV-1,HHV-2,HHV-3,HVT,MDV-1,MDV-2 and PRV were between 99.7%-11.9% and 99.4%-15% respectively.These data suggested that gE gene was conservative among different ILTV strains,and there existed very low homologous between different herpesvirus.
Key words:  infectious laryngotractheitis virus  Yantai strain  glycoprotein E gene  clone  sequence analysis