梅毒螺旋体Hsp40蛋白的生物信息学分析及基因克隆

Bioinformatical analysis and gene clone of treponema pallidum Hsp40

目的分析梅毒螺旋体(Treponema pallidum,Tp)Hsp40蛋白的生物学特性并克隆其基因。方法通过GenBank查到Hsp40蛋白的序列,利用生物信息学软件分析其生物学特性。根据Tp Hsp40基因序列设计特异性引物,利用PCR扩增Hsp40基因,并把该DNA片段连接到pMD19-T载体,转化大肠杆菌后挑取阳性克隆采用PCR和DNA测序进行验证。结果 Tp Hsp40基因能够编码374个氨基酸的蛋白质,其分子量为40.1 k Da,无信号肽,呈亲水性。Swissmodel预测得到Tp Hsp40的三级结构与嗜热杆菌(Thermus thermophilus)Hsp40结构最接近。从Tp基因组中扩增得到1125 bp的Tp Hsp40基因并将其连接到pMD19-T载体。结论本文预测了梅毒螺旋体毒力因子Hsp40蛋白的结构和克隆得到Tp Hsp40基因。

Objective To investigate the biological characteristics of treponema pallidum （Tp） Hsp 40 and clone its gene. Methods Protein sequence of Hsp40 was found using GenBank and its biological characteristics were analyzed using bioinformatical software. Specific primer was designed based on the genetic sequence of Tp Hsp40 and PCR was used to amplify the gene of Hsp40. The DNA fragment was ligated to pMD19-T carrier and transformed to Escherichia eoli. Positive clones were selected to be verified using PCR and DNA sequencing. Results Gene of Tp Hsp40 could en- code a protein of 374 amino acids. The molecular weight of the protein was 40.1 kDa. The protein had no signal peptide and was hydrophilie. The tertiary structure of Tp Hsp40 predicted by Swissmodel was closest to the structure of thermus thermophilus Hsp40. Gene of Tp Hsp40 with 1125bp amplified from genome of Tp was ligated to pMD19-T carrier. Conclusion This paper predicted the structure of Hsp40, the virulence factor of Tp, and cloned the gene of Tp Hsp40.