猪囊尾蚴RNA聚合酶亚基基因的筛选及结构初探

Screening of cDNA Clone for Putative RNA Polymerase Subunit of Cysticercus cellulosae

的 从构建的剪切引导序列 (SL)cDNA文库中筛选猪囊尾蚴相关基因。 方法 提取猪囊尾蚴总RNA ,利用SL特异序列和带有噬菌体M13M4的olig(dT)为引物 ,反转录成cDNA ,构建猪囊尾蚴SLcDNA文库。随机筛选并通过酶切、PCR鉴定阳性克隆 ,分析其同源性。 结果 鉴定出一 3 3 2bp的插入片段 ,含有 2 0 4bp的开放阅读框(ORF)和 3′端 2 0bp的polyA尾巴 ,经核苷酸和氨基酸序列分析 ,所克隆的基因的氨基酸序列与其他真核生物如人、秀丽隐杆线虫、果蝇及拟南芥等的RNA聚合酶亚基基因同源性均高达 71.6%以上。 结论 筛选鉴定的基因推测为猪囊尾蚴RNA聚合酶亚基基因 ,而且在不同物种间很保守。

Objective To obtain related genes of Cysticercus cellulosae from spliced leader (SL) cDNA library. Methods Spliced leader library of Cysticercus cellulosae was constructed using SL specific primer and oligo (dT)15 with M13M4 primer, and positive clones were then screened randomly, identified with enzyme restriction, followed by sequencing and homologous analysis. Results The amino acid sequence, encoded by the positive clone with a poly (A) 22 tail and a complete open reading frame (ORF), was with homology of RNA polymerase subunit genes of human, B. napus, fission yeast, A. thaliana, C. elegans and fruit fly up to 71.6%. Conclusion The protein, RNA polymerase subunit encoded putatively by the clone, is high conservative in different species.