title: [High-effective expression and construction of a prokaryote expression vector pET30a-glutathione-S-transferase A1 gene from human liver cDNA library and sequence determination].
abstract: 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.
authors.affiliation: Public Health School, Zhengzhou University, Zhengzhou 450052, China.
authors.forename: Pingping
authors.initials: P
authors.lastname: Chen
authors.forename: Chaowu
authors.initials: C
authors.lastname: Zhang
authors.forename: Yiming
authors.initials: Y
authors.lastname: Wu
authors.forename: Liangfeng
authors.initials: L
authors.lastname: Hang
authors.forename: Lingling
authors.initials: L
authors.lastname: Chen
mesh_terms: 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
chemical_list: D018076:DNA, Complementary; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase