Cloning of human PRL-3 gene and construction of its prokaryotic expression vector.
- Author:
Jun ZHOU
1
;
Jian-ming LI
;
Yu-hong LIU
;
Yan-qing DING
Author Information
- Publication Type:Journal Article
- MeSH: Base Sequence; Cell Line, Tumor; Cloning, Molecular; DNA, Complementary; chemistry; genetics; Electrophoresis, Polyacrylamide Gel; Escherichia coli; genetics; Genetic Vectors; genetics; Humans; Molecular Sequence Data; Neoplasm Proteins; genetics; metabolism; Protein Tyrosine Phosphatases; genetics; metabolism; Recombinant Fusion Proteins; genetics; metabolism; Sequence Analysis, DNA
- From: Journal of Southern Medical University 2007;27(5):641-643
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo obtain the entire coding sequence of human PRL-3 gene and construct its prokaryotic expression vector.
METHODSWith total RNA extracted from the human colorectal carcinoma cell line SW480 as the template, PRL-3 gene was amplified by RT-PCR with primers designed according to the published sequence of GenBank, and the product was inserted into pGEM-T Easy vector. The recombinant plasmid pGEM-T-PRL-3 was identified by restriction endonuclease analysis and DNA sequencing. After digestion with restriction endonuclease, PRL-3 gene was cloned into the multicloning sites of the prokaryotic expression vector pGEX-4T-1.
RESULTS AND CONCLUSIONThe entire coding region of human PRL-3 gene was cloned, and the recombinant pGEX-4T-1-PRL-3 vector was successfully constructed and expressed, which may provide the basis for further study of the relationship between human colorectal carcinoma and PRL-3 gene.
