Cloning of inorganic pyrophosphatase(PPA) gene and construction of its plant expression vetor

To use inorganic pyrophosphatase(PPa)regulating sucrose synthesization in transgenic sugarcane research,the PPa was cloned from Yeast.and its sequences were refered to Genbank and were confirmed by comparison with the known genome sequences of others.The sequence and bioinformatics analysis indicated that,the ORF sequence were 864 bp in length and encode 287 amino acid.By comparison,the homology of Nucleotide and amino acid were 99 % and 86 % respectively between the Saccharomyces cerevisiae,S.cerevisiae chromosome II,Yeast PPA gene and other isolates from Candida glabrata chromosome H,Candida glabrata partial mRNA.The analysis of hydrophilicity showed that it was hydrophilic.The cloned rbcS promoter,which the 5'-upstream regulation region of rice Rubisco small subunit gene(rbcS) was cloned from a Chinese cultivar Wuyunjing 8,was fused to the 5'-upstream of PPa gene coding region in a binary vector,and the rbcS-PPa gene was transmited into Agrobacterium tumefaciens.The integration of the rbcS-PPa fusion gene in Agrobacterium tumefaciens was confirmed by PCR analysis.