嗜热真菌Thermomyces langinosus编码丝/苏氨酸蛋白激酶部分cDNA的克隆及序列分析(英文)

Cloning and sequencing of a cDNA coding a putative ser/thr kinase from Thermomyces langinosus

根据几种丝状真菌保守的氨基酸序列,设计了一组简并性的引物,从嗜热真菌Thermomyceslanginosus中提取总RNA,利用RT-PCR的方法,我们得到了一个1243bp的cDNA的片段。片段回收纯化后连接到pGEM-Teasy载体上,PCR扩增重组质粒证实这个长约1200bp片段已经插到载体上。序列分析发现和裂殖酵母的蛋白激酶dis1非常相似,比较两者3'氨基酸序列发现同源性达28%以上。其推断的氨基酸序列上包含有6个丝/苏氨酸蛋白激酶的保守亚区。这些都表明它编码的基因可能是一个新的丝/苏氨酸蛋白激酶。

Based on the conserved amino acid sequence of ser/thr protein kinase from several filamentous fungus,degenerate primers was designed and synthesized.Total RNA was isolated from the fungi Thermomyces langinosus.Using RTPCR,one 1243bp cDNA fragment was obtained,and was cloned into pGEMT easy vector.PCR amplification indicated that the 1243bp fragment was inserted into the plasmid vector.Sequencing analysis showed that the cloned cDNA of Thermomyces langinosus was similar to that of fission yeast protein kinase disl.Comparison results showed that the homogenity of the 3'deduced amino acids was above 28%.Six conservative amino acid subdomains which most ser/thr kinases contain can be found in the deduced amino acid sequence.These showed that it might be a novel ser/thr protein kinase.