A series of novel directional cloning and expression vectors for blunt-end ligation of PCR products.
Novel directional cloning and expression vectors were developed for blunt-end ligation of PCR products that are suitable for high-throughput cloning and simplifying the screening procedure. The PCR products, without further processing, are cloned into vectors digested with SchI and, following transformation, the desired recombinants give typical blue colonies on selectable plates. The principle of this selection strategy is that the construction also generates a full-length ideal lacO gene. To the best of our knowledge, this is the first time that this lacO reconstruction strategy has been applied in the selection of recombinants.
Wu
Qiong
Q
Institute of Molecular Biology, Biology Faculty of Hubei University, Wuhan, Hubei Province, 430062, People's Republic of China.
Zhong
Xing
X
Zhai
Chao
C
Yang
Ju
J
Chen
Xiaomei
X
Chen
Liang
L
Wang
Wenfeng
W
Ma
Lixin
L
D003001:Cloning, Molecular / Q000379:methods*; D004926:Escherichia coli / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016366:Open Reading Frames / Q000235:genetics; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*