Cloning of Squalene Synthase Gene from Saccharomyces cerevisiae and Construction of Yeast Expression Vector / 广州中医药大学学报

ABSTRACT

Objective To lay the foundation for studying the synthesis of artemisinin in microorganism,squalene synthase(SS) gene,a key enzyme gene from Saccharomyces cerevisiae,was cloned and a yeast expression vector was constructed.Methods After amplification of SS gene by polymerase chain reaction(PCR),ligation to T-vector and analysis of the cloned sequence,enzyme digestion and reconfirmation of the target gene,the antisense yeast expression vector was constructed by inverted insertion of the target gene into a yeast expression vector,pGAPZ?A,and digested with two restriction enzymes for vertifying the recombinant.Results The length of SS gene was 1335bp.The preliminary sequence data indicated that SS gene obtained from the experiment had a high sequence homology with that from GenBank,except for a few base pairs.The antisense yeast expression vector has been constructed and vertified by digesting with two enzymes.Conclusion SS gene from Saccharomyces cerevisiae has been successfully cloned and sequenced.An antisense yeast expression vector has been also constructed.