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目的扩增结核分枝杆菌(H37Rv)的higB基因,运用生物信息学方法预测其编码蛋白的生物信息学特征。方法采用PCR方法扩增higB基因并对扩增产物测序,通过在线Protparam、protscale、SignaIP 4.1、MotifScan、SWISS_MODEL等程序,分析higB基因编码蛋白的生物学特征。结果获得的higB基因全长378bp,与预期大小基本一致,基因序列与GenBank上公布的MTB标准株的核苷酸同源性为100.00%。higB蛋白共125个氨基酸,分子质量单位为14.429 8ku,理论等电点10.18,脂溶性系数77.36,不稳定系数35.67,预测该蛋白为稳定蛋白。higB蛋白无信号肽,其二级结构中β-转角占12%,无规则卷曲占34.4%,氨基酸序列中有3个潜在的B细胞抗原表位,4个Th细胞表位,5个磷酸化位点。结论成功扩增了结核分枝杆菌higB基因,通过生物学信息获得了其编码蛋白的信息学特征,为研究该蛋白的生物学功能和免疫活性奠定了基础。
Objective To amplify the higB gene of Mycobacterium tuberculosis (H37Rv) and predict its bioinformatics characteristics by bioinformatics methods. Methods The higB gene was amplified by PCR and sequenced. The biological characteristics of higB gene were analyzed by on-line Protparam, protscale, SignaIP 4.1, MotifScan and SWISS_MODEL programs. The resulting higB gene was 378 bp in length, which was consistent with the expected size. The nucleotide sequence of the higB gene was 100.00% homologous to the MTB standard strain published in GenBank. The higB protein is 125 amino acids in total. The molecular mass unit is 14.429 8 ku, the theoretical isoelectric point is 10.18, the liposolubility coefficient is 77.36, and the instability coefficient is 35.67. The protein is predicted to be a stable protein. The higB protein has no signal peptide. Its secondary structure contains 12% β-turn, 34.4% random coil, 3 potential B-cell epitopes, 4 Th-cell epitopes and 5 phosphorylation in the amino acid sequence Site. Conclusion The higB gene of Mycobacterium tuberculosis was successfully amplified and the informative characteristics of its encoded protein were obtained through biological information, which laid the foundation for the study of the biological function and immunological activity of higB gene.