扩展莫尼茨绦虫蛋白激酶C相互作用蛋白(PICK1)基因的克隆及序列分析

Cloning and Sequence Analysis of the cDNA Encoding Protein Interacting with C Kinase 1 (PICK1) in Moniezia expansa

【目的】分离和鉴定扩展莫尼茨绦虫(Moniezia expansa)新基因,为进一步研究该基因的功能奠定基础。【方法】构建扩展莫尼茨绦虫成虫cDNA文库,随机挑取重组阳性克隆进行测序,对部分序列进行引物步移法测序,获取其全长cDNA序列;采用生物信息学等分析技术对该cDNA序列进行开放阅读框(ORF)的寻找、编码氨基酸的推导、核苷酸和氨基酸同源性比较及蛋白质二级结构的初步预测。【结果】获得了1个扩展莫尼茨绦虫新基因蛋白激酶C相互作用蛋白,全长1 527 bp,编码447个氨基酸,CDS预测存在明显的BAR,PDZ结构域。编码蛋白的理论分子质量为50.173 3 ku,等电点为5.22。【结论】获得了扩展莫尼茨绦虫蛋白激酶C相互作用蛋白的全长cDNA序列,为该基因功能的试验性鉴定工作奠定基础。

【Objective】The purpose of this program was to clone and identify novel genes from an adult Moniezia expansa（M.expansa） cDNA library,and provide a foundation for further research.【Method】A cDNA library was constructed from M.expansa adult stage.Clones were selected randomly from the cDNA library and were sequenced by using the method of expression sequence tags（ESTs）.Novel genes were acquired by primer-walking.The cDNA sequence encoding M.expansa PICK1 protein was analyzed,including searching the ORF,translating the nucleotide to protein sequence,similarity searches and secondary structure predication with bioinformatics analysis.【Result】PICK1 genes,1 527 bp and coding for 447 amino acids,was cloned and sequenced,then the sequence was submitted to GenBank and got an accession number,GH291479.The theoretical pI was 5.22 and molecular weight was 50.173 3 ku.【Conclusion】The full-length cDNA encoding M.expansa PICK1 was obtained,which laid the foundation for further functional study of this gene.