[Over-expression of glutathione S-transferase in Streptomyces]

mRNA was isolated and purified from human liver, and it was also used as templet for cDNA synthesis under the existence of reverse transcriptase. Two primers were designed and synthesized according to GST gene sequence which has been reported, GST gene was obtained using cDNA as templet and PCR technique. The sequencing result indicated that the GST gene is reliable, it was subcloned into NdeI and Bg1 II sites of plasmid pIJ6021, and then introduced into Streptomyces lividans TK54. Proteins were isolated from transformants (TK54/pIJ4486 and TK54/pIJ6021) respectively, SDS-PAGE result showed that the GST over-expressed and its yield is about 15% in soluble proteins in Streptomyces.