植物甜蛋白mabinlinⅡcDNA的克隆与序列分析

Cloning and Sequencing of cDNA Encoding Sweet tasting Plant Protein Mabinlin Ⅱ

根据已由作者克隆的ｍａｂｉｎｌｉｎⅡＢ亚基１８５ｂｐ的ｃＤＮＡ片段，设计合成系列引物．从马槟榔（ＣａｐｐａｒｉｓｍａｓａｉｋａｉＬéｖｌ．）种子提取总ＲＮＡ并纯化ｍＲＮＡ，经反转录合成ｃＤＮＡ第一链．采用ＲＡＣＥ方法，快捷克隆ｍａｂｉｎｌｉｎⅡ全长ｃＤＮＡ．经序列测定与分析，该ｃＤＮＡ为５８３ｂｐ，其中编码序列长４６５ｂｐ，共编码１５５个氨基酸．

A series of primers were synthesed according to the 185 bp of cDNA encoding B subunit of mabinlin Ⅱ cloned by the author based on its amino acid sequence.mRNA was extracted from the seeds of Capparis masaikai and purified.The full length cDNA of mabinlin Ⅱ was cloned by rapid amplification of cDNA ends(RACE).The result shows that the amplified DNA are 583 bp and its coding sequence is 465 bp and encodes 155 amino acids.