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A cosmid is a type of hybrid plasmid that contains a Lambda phage cos sequence . Cosmids (cos sites + plasmid = cosmids) DNA sequences are originally from the lambda phage. They are often used as a cloning vector in genetic engineering. Cosmids can be used to build genomic libraries. They were first described by Collins and Hohn in 1978. Cosmids can contain 37 to 52 (normally 45) kb of DNA, limits based on the normal bacteriophage packaging size. They can replicate as plasmids if they have a suitable origin of replication (ori): for example SV40 ori in mammalian cells, ColE1 ori for double-stranded DNA replication, or f1 ori for single-stranded DNA replication in prokaryotes. They frequently also contain a gene for selection such as antibiotic resistance, so that the transformed cells can be identified by plating on a medium containing the antibiotic. Those cells which did not take up the cosmid would be unable to grow.
A three-gene deleted bovine herpes virus 1, its construction method and application
This invention belongs to the field of viral genetic engineering technology and discloses a bovine herpesvirus type 1 (BHV-1) with three gene deletions, its construction method, and its applications. Using a Fosmid library, four myxovirus rescue combinations were screened. FosBHV-1-b-ΔTK-mCherry and FosBHV-1-d-ΔgEct / US9-eGFP were obtained through Red / ET and Gateway methods. These were then combined with FosBHV-1-a and FosBHV-1-c to construct an infectious cloning platform for recombinant BHV-1 with the TK / gEct / US9 three-gene deletions, rescuing recombinant viruses carrying tracer genes. This method has high recombination efficiency, and the cloning platform can efficiently construct BHV-1 gene-deleted strains. The immunogenicity of the deleted strains is comparable to that of wild-type viruses, with good safety, providing an efficient technical means for the development of BHV-1 attenuated vaccines and bovine disease live vector vaccines.
Owner:CHINA AGRI UNIV