[High-effective expression and construction of a prokaryote expression vector pET30a-glutathione-S-transferase A1 gene from human liver cDNA library and sequence determination].
OBJECTIVE
To express highly effective on IPTG-induced and construct an prokaryote expression vector carrying human glutathione-S- transferase (GST) A1 gene and to determine partial sequence analysis.


METHODS
The GSTA1 cDNA was amplified and extracted from human liver total RNAs by RT-PCR approach and recombined with prokaryote expression vector pET30a. The recombined vector pET30a-GSTA1 was verified using PCR, restriction analysis and sequencing determination and induced with IPTG in 35 degrees C .


RESULTS
Human GSTA1 gene was recombined corrected with pET30a, compared with Genbank , in code 512 T-->C, amino acid Met-->Thr; alignment core is 99% . The expressed fusion protein,with molecular weight of about 34.4 KDa, were about 41.8% (1h), 68% (2h), 68.3% (3h), 83% (4h) of the total cell protein by SDS-PAGE.


CONCLUSIONS
The protein expression of GSTA1 was demonstrated by Western blotting, it is correct.
Chen
Pingping
P
Public Health School, Zhengzhou University, Zhengzhou 450052, China.
Zhang
Chaowu
C
Wu
Yiming
Y
Hang
Liangfeng
L
Chen
Lingling
L
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics; D015870:Gene Expression; D015723:Gene Library; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D006801:Humans; D008099:Liver / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017385:Sequence Homology
D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase