Constructing recombinant herpesvirus BAC vectors with mating-assisted genetically integrated clone method.
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.
Jiang
Sijing
S
National Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, 430070, Hubei, People's Republic of China. jiangsijing@hubu.edu.cn
Zhong
Xing
X
Zhai
Chao
C
Chen
Liang
L
Ma
Lixin
L
Jin
Meilin
M
Chen
Huanchun
H
D022202:Chromosomes, Artificial, Bacterial; D003001:Cloning, Molecular; D004261:DNA Replication; D032621:DNA Shuffling; D004926:Escherichia coli / Q000235:genetics; D018390:Gene Targeting / Q000379:methods*; D005818:Genetic Engineering / Q000379:methods*; D005822:Genetic Vectors; D011558:Herpesvirus 1, Suid / Q000235:genetics*; D010957:Plasmids; D011995:Recombination, Genetic