Artificial chromosome transfer vector for recombinant herpesvirus-of-turkey bacteria

Abstract

The invention relates to an artificial chromosome transfer vector for recombinant herpesvirus-of-turkey bacteria, and belongs to the field of biological pharmacy. The artificial chromosome transfer vector for the recombinant herpesvirus-of-turkey bacteria is constructed by the following steps of: amplifying a xanthine-guanine phosphotransferase gene (gpt gene) in E.coli DH5alpha chromosome DNA, connecting the xanthine-gpt gene and a CMV promoter in plasmid pEGFP and poly (A), inserting the connected body into plasmid pUAB to construct recombinant clone plasmid pUAB-gpt with Egpt; and preparing a high-copy BAC vector basic functional sequence by using a pUC18 clone single-copy BAC vector, and inserting the high-copy BAC vector basic functional sequence into the pUAB-gpt to obtain the artificial chromosome transfer vector for the recombinant herpesvirus-of-turkey bacteria pUAB-gpt-BAC, wherein the pUAB-gpt-BAC contains a core sequence BAC-gpt of a gpt gene expression box and has double resistances of ampicillin and chloramphenicol. The constructed transfer vector has the application prospect for developing recombinant HVT live virus vector gene engineering vaccine.

Description

1. Technical field [0001] The invention relates to a recombinant turkey herpes virus bacterial artificial chromosome transfer carrier, in particular to the molecular design, technical route, transfer carrier and its application in the development of recombinant HVT live virus carrier genetic engineering vaccine, and belongs to the field of biopharmaceuticals. 2. Background technology [0002] Herpesvirus of turkey (Herpesvirus of turkey, HVT) is a non-pathogenic α-herpes virus, which was first isolated from turkeys in the late 1960s. It belongs to MDV-3 type, and its genome is linear and double-stranded. A DNA molecule, approximately 160kb in size. HVT is genetically and serologically related to Marek's disease virus (MDV), the causative agent of the highly contagious, neoplastic disease Marek's disease (MD) in chickens, and has therefore been used as a vaccine against MD since the 1970s strains are widely used around the world. In addition, HVT, as a viral vector carrying...

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Problems solved by technology

[0003] Due to the large size of the HVT genome, its complex structure, and its str

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