一种新的钙依赖性的蛋白激酶基因CP4的克隆及其表达研究

Cloning and Expression Analysis of a New Gene of CDPKs-CP4 from Arabidopsis

在利用人ACA (Anti Centromere/KinetochoreAutoantibody)血清研究拟南芥ACA相关蛋白的过程中克隆了钙依赖性的蛋白激酶基因家族的一个不典型基因 ,称为CP4 ,GenBank注册号为AF130 2 5 2。Southern杂交结果表明 ,拟南芥基因组中至少存在两种以上等位形式的CP4。原位杂交结果表明 ,CP4mRNA在拟南芥的花和根中高表达 ,在果实和叶片中没有表达。此外还构建了CP4的高效表达载体 ,为进一步研究CP4的功能打下了基础。

We cloned an atypical gene, designated CP4 (Accession number AF130252) which encodes a new, divergent isoform of this family, from Arabidopsis thaliana by screening a cDNA-expression library using ACA (anti-centromere/kinetochore autoantibody) serum. The squence of CP4shows 60% nucleotide identities with several other CDPKs. It contains four conserved serine/threonine kinase catalytic domains at N-terminal, but no conserved EF-hand motif in C-terminal, neither is the junction domain. Southern blot analysis detected that there are at least 2 kinds of CP4 isoforms in Arabidopsis. In situ hybridization experiments revealed strong CP4 expression in flower and root. We also expressed CP4 to study its function further.