Cloning of ScINO1 Gene and Construction of Yeast Multi-copy Integration Expression Vector

The inositol-1-phosphate synthase encoding gene(ScINO1) was amplified from the genomic DNA of Saccharomyces cerevisiae S288c by PCR.The length of ScINO1 gene was 1 602 bp.The Blast results showed that ScINO1 gene sequence was consistent with sequence of S.cerevisiae S288c in GenBank.The digested products of PGK,CYC1 and a rDNA fragment were inserted into S.cerevisiae shuttle plasmid PUG6 to generate rDNA mediated multi-copy integrated expression vectors pURH. Then,to obtain multi-copy integrated expression vectors pURIH,the ScINO1 gene was inserted into pURH.The yeast multi-copy integration expression vector was further verified through double restriction enzyme digestion.The construction of yeast multi-copy integration expression vector was provided with basic material for further study in genetic engineering of synthesis inositol and high ethanol tolerance in yeast.