通过RACE克隆得到了决明MCCase的cDNA,利用生物信息学进行系统分析,并利用荧光定量PCR对决明MCCase进行了差异表达分析。结果表明:决明MCCase全长1 269bp,编码423个氨基酸的蛋白质,为偏酸性不稳定蛋白质,定位于线粒体或细胞质中。决明MCCase与拟南芥的MCCase同源性最高。MCCase上有一个保守的功能结构域,与乙酰CoA羧化酶显示出较高的相似性,并在该结构域内发现了4个高度保守的氨基酸基序,可能在催化过程中发挥关键作用。MCCase在不同组织中存在差异表达,在根与胚轴中显示出高表达,在其它组织中表达量低,这可能与光和糖类抑制MCCase的表达有关。
The cDNA sequence of Cassia obtusifolia MCCase was abtained by RACE and analyzed by bioinformatics.The tissue-specific expression of MCCase was analyzed by quantitation RT-PCR.The results showed MCCase sequence had length of 1 269 bp and coded a protein consisting of 423 amine acids,which was an acid,hydrophilic,unstable protein and was located to mitochondria or cytoplasm.Cassia MCCase had the highest homology with Arabidopsis MCCase.There was a conserved functional domain in MCCase,which was similar to that of acetyl CoA carboxylase.Four highly conserved motifs were found in the conserved domain and might play important role in catalysis.MCCase expressed diferently in various tissues.It showed high expression level in root/hypocotyl and low expression level in other tissues.This might be related to the expressional inhibition of MCCase caused by carbohydrate and light.