ACTA VETERINARIA ET ZOOTECHNICA SINICA ›› 2014, Vol. 45 ›› Issue (8): 1309-1316.doi: 10.11843/j.issn.0366-6964.2014.08.015

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Preparation of Polyclonal Antibodies against gC Protein of Infectious Laryngotracheitis Virus，and Intracellular Localization and Function Research of gC Protein

SUN Yong-zhen1,2，ZHAO Yan1,2，CUI Hong-yu1,2，WANG Yun-feng1,2*   

  1. （1.State Key Laboratory of Veterinary Biotechnology/Avian Respiratory Disease and Immunology Group，Harbin Veterinary Research Institute，Chinese Academy of Agricultural Sciences，Harbin 150001，China；2.National Engineering Research Center of Veterinary Biologics，Harbin 150001，China）
  • Received:2014-01-09 Online:2014-08-23 Published:2014-08-23

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Abstract:

To prepare antibody with diagnosis value to infectious laryngotracheitis and to study the distribution of ILTV gC protein in LMH cells and the role of gC protein in the spread of ILTV among cells，the major antigen coded sequence of gC gene was amplified from ILTV-LJS09 genome by PCR.The prokaryotic expression vector pET-32a-gC was constructed by inserting the gC gene into pET-32a(+).The His-gC recombinant protein was purified，and then was used to immunize rabbits.The Western blot results demonstrate that the obtained polyclonal antibodies were specific to ILTV-LJS09 gC protein.The IFA results showed that the anti ILTV-LJS09 gC protein antibody could specifically response to ILTV-LJS09，HLJ0507，MDJ0442，SD0203，Zhj1298 strain.Intracellular localization of the ILTV gC protein in LMH cells using the anti-gC antibody showed that gC protein mainly located in cytoplasm and membrane of the virus infected cells.Antibody neutralization test indicated that the gC protein may be directly involved in virus transmission between the cells，thus affecting the virus plaque formation.In this study，we successfully prepared rabbit anti ILTV-LJS09 gC protein polyclonal antibody with diagnosis application value，and proved that LJS09 gC protein had the conservatism of α-herpesvirus on the distribution and function，which provided important data to the research of gC protein biological characteristics and ILTV infection mechanism.

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