[A universal high-throughput novel method of constructing the vectors].
In this paper, a simple universal high-throughput method of constructing the vectors was developed. It could add proper adapter when the PCR primers were designed, and the purpose fragments were cloned by PCR, and the various complementary sticky ends were created by T4 DNA polymerase's 3' -exodeoxyribonuclease activity. If all these fragments were put together with DNA ligase, they would recombinate in an orientation. If they had been transformated, the tansformants would be identified. Let's take the Oryza sativa single-cross homologous recombination chloroplast expression vector pRSMGA which was constructed with seven fragments as an example, if the vector pRSMGA was constructed in using the method what had mentioned, only twice recombination and transformation would be done. Scores of experiments had proved that it is a simple universal high-throughput novel method to construct the complicated vectors, which has not appeared in the periodical.
Lu
Yun-Hua
YH
Laboratory of Molecular Microbiology and Gene Engineering, College of Life Science, Hubei University, Wuhan 430062, China. lyh655@yahoo.com.cn
Ma
Li-Xin
LX
Jiang
Si-Jing
SJ
D005822:Genetic Vectors / Q000235:genetics*; D008957:Models, Genetic; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction