猪瘟病毒E2基因在昆虫细胞中的分泌表达及活性检测  

Expression of Secreted Forms of CSFV E2 Gene in Insect Cells and the Antigenic Activity of Recombinant E2 Protein

作　　者：查云峰[1] 徐兴然[1] 肖昌[1] 余兴龙[1] 涂长春[1] 

机构地区：[1]军事医学科学院军事兽医研究所,长春130062

出　　处：《畜牧兽医学报》2005年第10期1100-1105,共6页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家"863"项目资助(2001AA213141)

摘　　要：The glycoprotein E2 contains major antigenic determinants and involved in inducing neutralization antibodies.A truncated form of the CSFV glycoprotein E2 was expressed in baculovirus.The signal sequences of E2 gene were replaced with the honeybee melittin signal sequence,allowing efficient entrance into the secretory pathway in insect cell.In addition,the hydrophobic transmembrane anchors at the carboxyl termini of E2 proteins were removed to enable secretion rather than maintenance in the cellular membranes.Recombinant E2 protein was purified from the supernatant of infected cells by employing Ni2+ affinity chromatography.Protein purified was recognized by at least three anti-E2 pig sera and WH211 monoclonal antibody.WH211 monoclonal antibody couldn′t recognize the protein expressed in E.coli,indicating that it retain native structure.Thus,E2 expressed in insect cells can be used as a tool for diagnostic tests as well as obtaining material that could be suitable for X-ray crystallography studies.The glycoprotein E2 contains major antigenic determinants and involved in inducing neutralization antibodies. A truncated form of the CSFV glycoprotein E2 was expressed in baculovirus. The signal sequences of E2 gene were replaced with the honeybee melittin signal sequence, allowing efficient entrance into the secretory pathway in insect cell. In addition, the hydrophobic transmembrane anchors at the carboxyl termini of E2 proteins were removed to enable secretion rather than maintenance in the cellular membranes. Recombinant E2 protein was purified from the supernatant of infected cells by employing Ni^2＋ affinity chromatography. Protein purified was recognized by at least three anti-E2 pig sera and WH211 monoclonal antibody. WH211 monoclonal antibody couldn＇t recognize the protein expressed in E.coli, indicating that it retain native structure. Thus, E2 expressed in insect cells can be used as a tool for diagnostic tests as well as obtaining material that could be suitable for X-ray crystallography studies.

关 键 词：猪瘟病毒 E2基因 杆状病毒表达系统 分泌表达 

分 类 号：S858.285.3[农业科学—临床兽医学] Q786[农业科学—兽医学]