Method for constructing universal carrier of I type poultry Marek's virus vaccine strain gene engineering and application thereof

Abstract

The invention relates to a method for constructing a universal carrier of I type poultry Marek's virus vaccine strain gene engineering and an application thereof. The construction of the universal carrier (T-Us) mainly comprises the steps of: designing and synthesizing green fluorescence and neomycin gene (S-g-n), highly-efficient melting a report and a selection gene expressed as the universal carrier in the regulation of an SV40 early promoter, selecting an unnecessary area US10 copied by MDV as an insertion point of an external gene, simultaneously selecting the appropriate length of an isogenous arm (U) of the universal carrier, left and right lengths being 1000-2000 bp respectively; modifying multi-colon sites by new synthesis, introducing a proper intron, constructing an expression box (C-M-A), and constructing the universal carrier (T-Us) after connecting all the elements. The universal carrier is cotransfected with the I type poultry Marek's virus to obtain a recombined I typepoultry Marek's virus vaccine CVI988-26 strain, thus establishing a firm foundation for developing new Marek's disease vaccine.

Description

technical field [0001] The invention relates to a method for constructing a general vector for genetic engineering of poultry type I Marek's disease virus vaccine strain and an application in the obtained recombinant poultry type I Marek's disease virus vaccine strain prescription, belonging to the field of veterinary biological products. Background technique [0002] Marek's Disease Virus (MDV), formally known as avian herpes virus type 2 (Gallidherpesvirus 2), is an important infectious disease pathogen of chickens and has tumorigenic properties (Edited by Lu Chengping. Veterinary Microbiology. Fourth Edition, China Agricultural Press, 2007, 359). Marek's Disease Virus (MDV) causes a highly contagious neoplastic disease in chickens, characterized by lymphoid cell infiltration, hyperplasia, and tumor formation in peripheral nerves, iris, skin, muscles, and various internal organs. Fast speed and long incubation period. It is prevalent worldwide and is one of the three maj...

AI Technical Summary

Problems solved by technology

[0005] In the MDV transfer vectors that have been reported now, the screening gene and the marker gene are used alone, which causes some problems and shortcomings in the application of the existing MDV transfer vectors.

If the screening gene is used alone, such as ampicillin and neomycin marker gene, its disadvantage is that it cannot visually display the quantity and purity of the recombinant virus, resulting in the blindness of the experiment

If the marker gene is used alone, such as Li Yongqing; Zhou Xuemei; Zhang Peijun, etc., the marker gene used alone is LacZ. The disadvantage is that during the screening process of recombinant viruses, the contamination of maternal MDV cannot be suppressed, resulting in the inability to effectively distinguish recombinant viruses from maternal MDV, which affects screening. Efficiency, resulting in long screening time and easy failure

On the other hand, existing vectors can only allow the insertion and expression of an exogenous antigen gene ("Recombinant Chicken Marek's Disease Virus Transfer Vector and Application" (publication number: CN1763205; )

The above shortcomings limit the application of existing transfer vectors, and cannot quickly construct recombinant viruses, hindering the production and application of recombinant MDV

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