Cloning and Sequence Analysis of α-Pinene Synthase Gene from Pinus massoniana
[Objective] To clone the full-length of α-pinene synthase gene from Pinus massoniana.[Method] According to high conservative amino acids of reported α-pinene synthase gene from pinaceaes,a pair of degenerate primers was designed and partial fragment was obtained by PCR amplification.Based on this known sequence,some new primers were designed,two terminals gene sequence were cloned by 3'RACE and 5' RACE respectively.After splicing the fragments of sequence,analyzed the structures and function of the coded protein by bioinformatics softwares.[Result] The full-length cDNA of α-pinene synthase gene consists of 2 103 bp,contains a 1 890 bp open reading frame (ORF) encoding 629 amino acid proteins,including N-terminal transit peptide,metal-binging domain and DDXXD motif.[Conclusion] The full-length cDNA of α-pinene synthase gene was cloned,which has the typical characteristics of monoterpene synthase gene.The sequence has been submitted to GeneBank,accessin No.KF547035.
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