大叶落地生根细胞周期蛋白依赖激酶KdCDK基因克隆及表达分析

c DNA Cloning and Expression Analysis of Cyclin-dependent Kinase(KdCDK) Gene in Kalanchoe daigremontiana

为了更好理解大叶落地生根中细胞周期调控的分子机制,利用RACE-PCR技术从大叶落地生根中克隆了1个新的基因KdCDK。该基因编码295个氨基酸残基,分子量为34.12kDa,等电点为6.15,其开放阅读框长度为888bp。其蛋白与笋瓜CDK蛋白的同源性最高,属于CDKA类蛋白家族。实时荧光定量PCR分析表明,该蛋白基因在大叶落地生根的根中表达量最高,且受渗透胁迫(甘露醇)的诱导上调表达。从大叶落地生根中克隆出KdCDK基因,为将来该基因的功能研究打下了基础。

To better understand the molecular mechanisms of CDKs involved in Kalanchoe daigremontiana,an A-Type CDK gene, KdCD K,was identified using rapid amplification of cDNA end（RACE）PCR. KdCDK gene consists of an ORF of 888 bp that was predicted to encode a 295 amino acid residue-long protein of 34.12 kDa with an isoelectric point of 6.15.KdCDK was related most closely to CmCDK and was clustered to CDKA subfamily.Real-time PCR analysis revealed that KdCDK transcript was expressed highly in root and upregulated under osmotic stress.This study characterized the novel KdCDK gene from Kalanchoe daigremontiana for the first time and the results would be useful for further functional determination of the gene.