中文摘要：

为了研究低等真核生物中蛋白激酶PKA（cAMP-dependent protein kinase A）的生物学功能,以八肋游仆虫（Euplotes octocarinatus）为实验材料,通过生物信息学方法分析八肋游仆虫基因组及转录组数据,发现八肋游仆虫中编码蛋白激酶PKAc（PKA催化亚基）的基因只有一种。通过PCR扩增获得游仆虫PKAc基因（EoPKAc）,分析表明EoPKAc基因全长1158bp,不含有内含子,开放阅读框981bp,编码326个氨基酸。序列比对结果显示EoPKAc含有与其它已知蛋白激酶相同的11个亚结构域（subdomain）以及一些重要的保守氨基酸。构建原核表达质粒pGEX-6P-1-PKAc,转化至大肠杆菌BL21中,EoPKAc获得高效表达,经GST柱亲和层析后获得较高纯度的EoPKAc蛋白,表达产物经Western blotting检测为阳性。

英文摘要：

To study the biological function of PKA（cAMP dependent protein kinase A）in lower eukaryotic organisms,the PKAgene from Euplotes octocarinatus was isolated and analyzed.Only one gene encoded PKAc（PKA C subunit）was identified by bioinformatic analysis of transcriptome and genome data from E.octocarinatus.The EoPKAc gene was obtained by PCR.The full length of EoPKAc was 1158 bp,which has no intron and contains an ORF of 981 bp encoding aprotein of 326 amino acids.The amino acid sequence analysis of EoPKAc showed the presence of all 11 subdomains and the conserved amino acids as other species.Recombinant plasmid pGEX-6P-1-PKAc was constructed and expressed in E.coli BL21.After purifying by GST affinity,the expressed products were checked by Western Blotting assay.