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目的对红色蚋细胞色素氧化酶亚基I(COI)基因进行克隆、鉴定与序列分析。方法以红色蚋成虫基因组DNA为模板,设计特异性引物进行PCR扩增、纯化基因片段后连接、转化大肠杆菌DH5a,筛选阳性克隆、测序并与NCBI数据库中的核酸序列进行同源性比对分析,掌握其基因序列及其蛋白质结构特征。结果克隆序列的结果表明COI基因全长1542 bp,编码513个氨基酸,编码蛋白等电点为5.84,相对分子量为56 KDa,具有一个细胞色素氧化酶亚基I的核心保守结构;其基因碱基组成A、T、C、G分别为28.60%,36.90%,17.76%,16.74%,G+C含量为34.5%,A+T含量为65.5%。结论成功克隆出红色蚋CO I基因序列,可为进一步研究奠定基础。
Objective To clone, identify and sequence the COI gene of red goblet cells. Methods The specific primers were designed and used to amplify the genomic DNA of the adult red moth, and then the gene fragments were ligated and transformed into E. coli DH5a. The positive clones were screened and sequenced. The sequences were compared with those of the NCBI database , Master its gene sequence and its protein structure characteristics. Results The cloned sequence showed that COI gene was 1542 bp in length and encoded a 513 amino acid protein with a pI of 5.84 and a relative molecular weight of 56 kDa. The COI gene has a core conserved structure of cytochrome oxidase subunit I. The composition A, T, C and G were 28.60%, 36.90%, 17.76% and 16.74% respectively, the G + C content was 34.5% and the A + T content was 65.5%. Conclusion The successful cloning of the COI gene sequence of red chrysin can lay a foundation for further research.