Fowl pox virus double-gene expression carrier (PT7.5N)

Abstract

The invention provides a fowlpox virus double-gene expression vector, which has a nucleotide sequence that is shown by SEQ ID NO.1. The invention includes a promoter P7.5, a LacZ gene, multiple cloning sites which are located on the LacZ gene, a rare restriction enzyme cutting site Not1, homologous arms TK1 and TK2 of a fowlpox virus DNA, a Kan <R> resistance marker gene, an origin of replication(ori) and a connection section. The double-gene expression vector constructed by the invention can reduce the length of the vector to the maximum extent, thus a larger exogenous gene can be contained; the number of available restriction enzyme cutting sites is increased so as to facilitate the cloning of different exogenous genes; the promoter at a insertion site Not1 is not limited and the promoter which is matched with the exogenous gene can be freely chosen so as to improve expression efficiency; furthermore, the invention adopts a reverse expression method so as to improve the expression efficiency of the exogenous gene.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a fowlpox virus double-gene expression vector, its construction method and application. Background technique [0002] Poxviridae is a large group of obligate viruses with a diameter of 300-400nm and a lipid-protein shell surrounding a complex core structure. This structure contains a linear nearly 20kb double-stranded DNA molecule, encoding multiple subunits, and DNA-dependent The RNA polymerase, transcription factor, cap structure and methylase, poly A polymerase, are all packaged in the viral core and translated into mRNAs. Fowlpox Virus (FPV) belongs to the genus Fowlpoxvirus of the family Poxviridae. The virion core contains double-strand linear DNA, G+C=35%, and the genome is about 300kb long. Under natural conditions, it can only infect poultry, but it can produce abortive infection on mammalian cells, does not produce infectious virus particles, but can express a...

AI Technical Summary

Problems solved by technology

[0003] At present, the existing fowlpox virus vectors are not perfect, such as the vector is too large, the length of the foreign gene that can be carried is limited, there are not many available restriction sites, and the operation is cumbersome

Moreover, most of the double-gene expression vectors are expressed in cis, which has a certain reduction in the expression of the second gene.

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