美洲大蠊泛素基因的克隆及生物信息学分析  

Cloning and bioinformatics analysis of an ubiquitin gene of Periplaneta americana

作　　者：金小宝[1] 刘雷山[1] 朱家勇[1] 马艳[1] 王艳[1] 

机构地区：[1]广东药学院基础学院寄生虫学教研室,广东广州510006

出　　处：《中国病原生物学杂志》2007年第5期371-373,392,共4页Journal of Pathogen Biology

摘　　要：目的克隆美洲大蠊(Periplaneta americana)泛素基因,并对序列进行相关分析。方法依据其他物种泛素基因的保守区设计1对简并引物,对美洲大蠊cDNA模板进行RT-PCR扩增,产物纯化后行T-A克隆、测序和酶切鉴定,运用相关生物信息学软件对该基因核苷酸序列进行分析和预测其编码蛋白的理化性质及二级结构等。结果从美洲大蠊体中克隆了泛素基因的编码区,被GenBank收录,登录号为EF101563。生物信息学分析结果表明,美洲大蠊泛素基因编码区的长度为228 bp,编码的多肽由76个氨基酸残基组成,相对分子质量单位为8.53 ku,理论等电点为6.56,与其他真核生物泛素基因在氨基酸水平上具有96%以上的相似性,二级结构中α-螺旋占26.3%,无规则卷曲占47.3%,延伸串占26.3%。结论成功克隆了美洲大蠊泛素基因,并进行了相关生物信息学分析,为进一步研究泛素在美洲大蠊机体内的作用奠定了基础。Objective To clone and analyze the Periplaneta americana ubiquitin gene. Methods The coding sequence of P. americana ubiquitin gene was amplified by RT-PCR with degenerated primers. Target PCR product was purified, cloned, sequenced and identified. Meanwhile, bioinformatics analyzed and predicted the physical-chemical characteristics and secondary structure of the gene. Results The length of this gene opening reading frame （ORF） was 228 bp, GeneBank Accession No. EF101563, encoding a protein with molecular weight of 8.54 ku and theoretical isoelectric point of 6.56. Multiple sequence alignment indicated that P. americana ubiquitin was very similar to the homologous proteins of other eukaryotic species and shared more than 96% identity with other eukaryotic ubiquitins at amino acid level. There are 26.3% α-helix, 47.3% random coil, 26.3% extended strand in the probable secondary structure of P. americana ubiquitin. Conclusion The P. americana ubiquitin gene is successfully cloned and analyzed, which is a basis to study on the function of ubiquitin in P. americana.

关 键 词：美洲大蠊 泛素基因 克隆 生物信息学 分析 

分 类 号：R384.9[医药卫生—医学寄生虫学]