马槟榔MBLⅡ基因重组及果实特异表达载体构建

Recombination of MBLⅡ gene and the construction of plant fruit-specific expressed vector

根据马槟榔(MBLⅡ)甜蛋白核酸序列及相应氨基酸序列的结构及功能预测,采用基因重迭延伸技术在编码A链和B链的序列间插入连接多肽LP4/2A,对MBLⅡ基因进行亚克隆并构建重组体.结果表明:试验克隆全长乙烯应答果实特异性E8启动子,构建了含除草剂抗性基因bar的E8启动子驱动的重组MBLⅡ基因植物表达载体pCA-E8-Mab,为实现甜蛋白基因在人参果果实中特异表达及改善风味奠定了基础.

According to the structure and function prediction of nucleic acid sequences and amino acid sequence of MBLⅡ,the peptide LP4/2A was inserted between encoding A chain and B chain sequences by adopting gene splicing-by-overlap extension（SOE） technique,then the MBLⅡ gene was subcloned and recombinant was constructed.The results showed the full-length fruit-specific promoter E8 with the sequence of ethylene responsiveness was cloned.The plant expression vector of pCA-E8-Mab with the herbicide resistance gene bar was constructed by the reconstructed MBLⅡ gene which drived by E8 promoter.This experiment built up a foundation for the following study of expressing sweet proteins gene in the pepino fruit and to improve its flavor.