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  • 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].
    Author: Chen P, Zhang C, Wu Y, Hang L, Chen L.
    Journal: Wei Sheng Yan Jiu; 2004 Sep; 33(5):596-9. PubMed ID: 15612490.
    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. CONCLUSION: The protein expression of GSTA1 was demonstrated by Western blotting, it is correct.
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