Yeast Surface Display Of Subgroup J Avian Multi-epitope Chimeric Gene Using Saccharomyces Cerevisiae

ALV(Avian Leukosis, AL) refers to the avian hematopoietic tissue overgrowth of certain cellular components belong mainly to the various subjects by retroviral retrovirus avian influenza retrovirus-induced infectious neoplastic disease, is a worldwide distribution of avian tumor diseases, one of the important disease has seriously hampered the development of the poultry industry. Since its discovery in 1911, ALV(Avian Leukosis Virus) study the pathogenesis of cancer has become an important model for sexually transmitted diseases, for its research scientists from various countries has been seriously. Since the virus covers a variety of subsets, there are different strains of the same subgroup, and associated widespread endogenous virus, resulting in the prevention and control of this disease greatly increased the difficulty. Western countries due to the advantages of breeding patterns and other aspects of management, through decades of efforts on large-scale farms infected ALV basic realization of purification. However, in recent years, ALV-J(J subgroup ALV) the occurrence and spread in our country has intensified the situation presented, there have been reports of ALV-J infection and local breeder flocks of white feather happen, ALV-J infection The incidence of early age also showed a trend, and for the first time found naturally infected patients with acute tumorigenic ALV-J trigger acute fibro-sarcoma. ALV-J on the development of the poultry industry has caused a serious threat to carry out large-scale farms for ALV purification has become an important task of China’s poultry industry, however, due to the geopolitical vast range chickens strains, culture conditions uneven and other reasons, long way to go to achieve ALV comprehensive purification. Many scholars try ALV vaccine development from the perspective of molecular biology to reduce ALV infection in chickens,in order to accelerate the pace of ALV purification.Multi-epitope vaccine(Multi-epitope Vaccine) is the use of molecular biology and computer science analysis, screening, access to the target antigen epitope outstanding, ideal for multiple epitopes optimized and developed a series of new vaccines. This vaccine is particularly suitable for gene mutation or the presence of oncogene easily genome of viruses such as human immunodeficiency virus(HIV), avian leukosis virus(ALV). By analyzing and get antigenicity of these viruses and conserved epitopes than the strong development of multiepitope vaccine is an important direction to overcome these tumor diseases.Yeast surface display system(Saccharomyces Cerevisiae Surface Display System) as a eukaryotic expression system, more and more attention in the antigen-antibody screening,protein directed evolution, water purification, and aspects of live vector vaccine has been widely used. Compared to E. coli, salmonella, etc., Saccharomyces cerevisiae is recognized as safe and nontoxic(GRAS, Generally Recognized as Safe) food grade industrial bacteria,widely used in food fermentation, animal feed and pharmaceutical engineering and other fields, the eukaryotic yeast post-translational modification of the expression system more advantages than bacteria.This study is the use of yeast surface display system and a multi-epitope vaccine-related technology, has been published by ALV-J virus structure on findings and bioinformatics software selected ALV-J and conserved antigenic good table alleles Gag(278-376aa), Pol(784-855aa), Env(Gp85: 145-156 aa and Gp37: 412-538aa) fragment series of multi-epitope gene X, use EBY100 / pCTCON2 yeast surface display system was constructed ALV-J multiepitope chimeric gene of Saccharomyces cerevisiae surface display vector EBY100 /pCTCON2-X, and multi-epitope chimeric gene X in the yeast cell surface expression were detected.The results show that the study successfully constructed ALV-J multi-epitope chimeric gene yeast surface display vector, after induction of expression in the yeast cell surface protein ALV- successfully demonstrated multi-epitope chimeric genes. Through the optimization of induction conditions, we found EBY100 / pCTCON2-X after the induction of the first 24 h, ALV-J multi-epitope chimeric gene expression levels of the highest expression efficiency of about 35%, up to 50%.In this study of ALV-J multi-epitope chimeric gene of Saccharomyces cerevisiae yeast surface display live vector vaccine study of ALV-J for the carrier for the subsequent foundation.