The temperature--dependent expression of GST of Schistosoma japonicum (Philippine strain)
Cai, Z.H.; Song, G.C.; Xu, Y.X.; Liu, S.X.
Southeast Asian Journal of Tropical Medicine and Public Health 24(1): 57-60
1993
ISSN/ISBN: 0125-1562 PMID: 8362308 Document Number: 413634
Obtained from pSj5, the cDNA gene encoding GST antigen of Schistosoma japonicum (Philippine strain) was ligated with efficient temperature-dependent PBV220 vector which was constructed in CAPM, and then introduced into host bacterium-DH5-alpha (Escherichia coli) by transformation. Transformants were selected by ampicillin and recombinant clones were identified by restriction mapping. The result showed that recombinant clone 43 was the one carrying recombinant plasmid PBV 220 with the correct insertion of the gene fragment. The GST expression ability of clone 43 was investigated by GST enzymic activity assay and SDS-PAGE. A relatively high level of GST enzymic activity was expressed by this clone under the temperature-dependent condition, that is, cultured at 30 degree C and expressed at 42 degree C. A more strongly stained 26 kDa protein band was identified by SDS-PAGE. The result indicated that GST of S. japonicum (Philippine strain) could be expressed not only by IPTG induction, but also by the temperature-dependent method.