中间锦鸡儿CiPP2C8-like基因的克隆及表达分析

Cloning and Expression Analysis of CiPP2C8-like Gene from Caragana intermedia

蛋白磷酸酶PP2C作为调节因子直接或间接调控逆境胁迫信号途径和生长发育过程。本研究从已构建好的抑制性消减文库(SSH)中获得1条PP2C基因序列,对其进行了克隆。测序表明该基因c DNA长为999 bp,共编码333个氨基酸,推导该蛋白分子量为36.12 k D,等电点为6.71,是一种亲水蛋白。经氨基酸序列比对和进化树分析表明,该基因属于PP2C家族成员,并命名为Ci PP2C8-like。实时荧光定量PCR检测表明中间锦鸡儿Ci PP2C8-like基因表达受盐和脱水胁迫诱导。

As a regulator,protein phosphatase PP2C directly or indirectly regulates stressing signaling pathways and the process of growth and development.In this study,a PP2C gene fragment was isolated from the ready-built suppression subtractive hybridization library（SSH） of Caragana intermedia and the full length c DNA was cloned.Sequencing results showed that the full length cDNA was 999 bp,encoding 333 amino acids with a calculated molecular mass of 36.12 kD and a theoretical pI of 6.71.The deduced protein was strongly hydrophilic.Amino acid sequence alignment and phylogenetic tree analysis showed that this gene belonged to the PP2C family,which was named as CiPP2C8-like.Real-time quantitative PCR analysis indicated that expression of CiPP2C8-like gene from Caragana intermedia was induced by the salt and dehydration stress.