| 研究简报 |
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| Gene Coloning, Expression and Enzymatic Assay of Human sPLA2-IIA |
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Abstract Objective To clone the cDNA of human sPLA2-IIA, construct the engineered Escherischia coli expressing human sPLA2-IIA and Identify the expressed human sPLA2-IIA. Methods: Total RNAs were purified from human fetal spleen. The cDNA of human sPLA2-IIA was cloned by RT-PCR and inserted into plasmid pET32a(+) between NcoI and EcoRI sites for expressing the recombinant human sPLA2-IIA in Escherischia coli BL21(DE3). The recombinants were screened by SDS-PAGE. The engineered Escherischia coli expressing trxA-human sPLA2-IIA fusion protein was established. The expressed human sPLA2-IIA exists in the form of inclusion body and accounts for about 25% of the total soluble proteins of Escherischia coli BL21(DE3). Conclusion: the engineered E. coli methods are suitable for preparing plenty of human sPLA2-IIA which has laid base for the large-scale expression, purification and basic studies of human sPLA2-IIA.
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Received: 07 April 2006
Published: 25 August 2006
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