关键词: 桑树, 查尔酮异构酶, 基因克隆, 组织表达, 原核表达

Abstract: The MaCHI gene was cloned from the ripe fruit of Morus alba 'Jialing 30’ by homologous cloning and suppression PCR. The full-length genomic sequence of MaCHI is 2 402 bp in length, with the 3'UTR of 290 bp and the open reading frame (ORF) of 2 112 bp. Its ORF consists of four exons and three introns, and its cDNA encodes 219 amino acids. The putative molecular weight of the protein encoded by MaCHI gene is 23.8 ku and its theoretical isoelectric point is 5.29. The RT-PCR was used to analyze the expression levels of MaCHI in different tissues, and its expression level was high in leaves and fruits, and relatively low in roots. A prokaryotic expression recombinant plasmid, pET-28a (+)-MaCHI was constructed and transformed into Escherichia coli for expression. This study will help establish theoretical foundation for the further research of MaCHI.

Key words: Morus alba, chalcone isomerase, gene cloning, tissue expression, prokaryotic expression