Constructing recombinant herpesvirus BAC vectors with mating-assisted genetically integrated clone method

Abstract

The large capacity of pseudorabies virus (PRV) for foreign DNA and broad host range make it a prospective tool for the preparation of vaccines and agents of gene and tumour therapy. Here we introduced a cloning strategy that facilitates construction of recombinant PRV-BAC vectors based on mating-assisted genetically integrated clone (MAGIC). The target gene was cloned into a small conditionally replicating donor plasmid, followed by shuffling to a recipient PRV-BAC plasmid in vivo of Escherichia coli through MAGIC. The average efficiency of successful clones was 89%. Moreover, permanent integration of unwanted sequences was avoided.

A novel approach for the construction of plant amiRNA expression vectors

“…It exploits bacterial mating, in vivo site-specific endonuclease cleavage and homologous recombination to catalyze the transfer of a DNA fragment between a donor vector in one bacterial strain and a recipient plasmid in another bacterial strain. But it has only been used for the construction of recombinant virus (Tan et al, 2006;Jiang et al, 2010). As almost all plant binary plasmids are more than 10 kb, it is difficult to apply conventional cloning process for plasmid construction.…”