马疱疹病毒1型主要毒力基因分析及TK基因缺失载体的构建

doi:10.16431/j.cnki.1671-7236.2018.10.004

Abstract

Abstract:

In order to understand the genetic evolution of the major virulence genes of equine herpesvirus 1 (EHV-1) of Xinjiang (XJ2015 strain) and construct TK gene-deleted strain,we cloned,sequenced and bioinformatics analyzed the full-length of TK,gI and gE genes of XJ2015 strain in this study.The DNA of the XJ2015 strain was used as a template,the TK gene recombination arms TKL and TKR were amplified to construct plasmid pUC-TKLR,and then inserted the amplified EGFP (CMV+polyA) into the pUC-TKLR plasmid.The results showed that it had higher homology between TK,gI and gE genes of XJ2015 strain and foreign EHV-1 isolates,which were 99.8% to 100.0%,99.6% to 100.0% and 99.9% to 100.0%,respectively,and had the lowest homology with foreign EHV-3 isolate,which were 72.9%,59.4% and 62.1%,respectively;Genetic evolutionary analysis showed that all three genes were genetically related to foreign EHV-1 isolates,the evolutionary relationship with EHV-3 was far away,but it was closely related to EHV-9 and EHV-4.There were no significant difference,no obvious regional features,the functional genes were conserved,the evolution was slow and homologous genes had the same or similar functions of TK,gI and gE genes of XJ2015 strain between foreign EHV-1 strains.After enzyme digestion,sequencing and transfection,the plasmid pUC-TKLR-EGFP for TK gene deletion targeting was successfully constructed.Through the analysis of EHV-1 main virulence genes and the construction of TK gene deletion targeting vector,it provided a theoretical basis for the epidemiological investigation and analysis of the TK gene deletion strain in Xinjiang.

Key words: equine herpesvirus type 1 (EHV-1); sequence analysis; gene deletion vector

CLC Number:

S852.65⁺2

FAN Bin, CHEN Zhuo, LIU Jianhua, BAO Zilei, HU Yue, JIA Qinrui, WANG Xuezhu, RAN Duoliang. Analysis of Major Viral Genes of Equine Herpesvirus Type 1 and Construction of TK Gene Deletion Vector[J]. , 2018, 45(10): 2681-2690.

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Analysis of Major Viral Genes of Equine Herpesvirus Type 1 and Construction of TK Gene Deletion Vector

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