巴西橡胶树HbCMK基因的克隆及表达

Cloning and Expression of HbCMK from Hevea brasiliensis

根据植物CMK(4-Diphosphocytidyl-2C-methyl-D-erythritol Kinase,CMK)的同源序列设计引物,通过RT-PCR结合RACE的方法在橡胶树中获得了与其相应的CMK基因,命名为HbCMK.序列分析表明HbCMK长1415bp,编码388个氨基酸,该氨基酸序列与长春花(Catharanthus roseus)、拟南芥(Arabidopsis thaliana)、甜菊(Stevia rebaudiana)、丹参(Salvia miltiorrhiza)、烟草(Nicotiana benthamiana)和水稻(Oryza sativa)的CMK相似性分别达到72.6%、72.5%、71.9%、70.9%、69.0%和66.9%.半定量RT-PCR结果显示,HbCMK的表达具有组织差异性,在愈伤组织中大量表达,在叶片和胶乳中微量表达,乙烯诱导胶乳HbCMK的表达,伤害对HbCMK的表达几乎没有影响.本实验结果为进一步了解MEP途径在橡胶树胶乳中的作用和天然橡胶生物合成的调控奠定了基础.

A 4-Diphosphocytidyl-2C-methyl-D-erythritol Kinase(CMK)gene was isolated from Hevea brasiliensis by RT-PCR and RACE based on the conserved sequences of plant CMK genes.The cDNA,which designated HbCMK,consists of 1 415 bp and encodes a protein of 388 amino acids with a predicted molecular mass of 43.19 kD.In comparison,this putative HbCMK protein shows from 66.9% to 72.6 % identity to other plants CMK.Semi-quantitative RT-PCR analysis indicates that the HbCMK is more highly expressed in callus than in latex ...