【摘要】：目的應(yīng)用生物學(xué)信息分析軟件預(yù)測(cè)結(jié)核分枝桿菌CarD蛋白的結(jié)構(gòu)和功能。方法運(yùn)用NCBI中Blast工具,將結(jié)核分枝桿菌H37Rv carD基因序列與GenBank中相似序列進(jìn)行核苷酸比對(duì);利用MEGA 6.06軟件,采用鄰位相連法構(gòu)建N-J進(jìn)化樹(shù);運(yùn)用ClustalX 2.1軟件,將結(jié)核分枝桿菌H37Rv CarD及與其同源性較高的分枝桿菌CarD氨基酸序列進(jìn)行多重比對(duì)分析;登錄ExPASy網(wǎng)站,運(yùn)用ProtParam工具分析CarD蛋白的理化性質(zhì),運(yùn)用ProtScale進(jìn)行蛋白質(zhì)疏水性分析,運(yùn)用SignalP 4.1Server預(yù)測(cè)蛋白質(zhì)信號(hào)肽,運(yùn)用TMHMM Server v.2.0以及TMPred預(yù)測(cè)蛋白質(zhì)跨膜螺旋區(qū),運(yùn)用SOPMA預(yù)測(cè)蛋白質(zhì)二級(jí)結(jié)構(gòu),運(yùn)用SWISS-MODEL進(jìn)行蛋白質(zhì)三級(jí)結(jié)構(gòu)同源建模;運(yùn)用NetPhos 3.1Server預(yù)測(cè)蛋白質(zhì)磷酸化位點(diǎn);運(yùn)用NCBI中CDD工具預(yù)測(cè)CarD蛋白保守結(jié)構(gòu)域。結(jié)果分枝桿菌carD基因序列相似度為100%,進(jìn)化關(guān)系較近。H37Rv與田鼠分枝桿菌起源于同一物種,同源性較高。CarD分子在進(jìn)化過(guò)程中高度保守,為穩(wěn)定、親水性蛋白,無(wú)跨膜區(qū)、無(wú)信號(hào)肽,蛋白序列中存在4個(gè)絲氨酸磷酸化位點(diǎn),二級(jí)結(jié)構(gòu)中以α-螺旋為主,三級(jí)結(jié)構(gòu)同源建模成功。CarD蛋白是一種結(jié)合RNA聚合酶的轉(zhuǎn)錄調(diào)控因子,屬于CarD/CdnL/TRCF家族。結(jié)論 CarD蛋白為RNA聚合酶結(jié)合蛋白,調(diào)控轉(zhuǎn)錄和控制結(jié)核分枝桿菌的生長(zhǎng)。該蛋白序列保守穩(wěn)定,是治療結(jié)核病潛在的新靶標(biāo)。
[Abstract]:Objective to predict the structure and function of Mycobacterium tuberculosis CarD protein by using biological information analysis software. Methods the sequence of H37Rv carD gene of Mycobacterium tuberculosis was compared with the similar sequence of GenBank by Blast tool in NCBI, and the N-J evolutionary tree was constructed by using MEGA 6.06 software. The amino acid sequences of Mycobacterium tuberculosis (H37Rv CarD) and Mycobacterium tuberculosis (CarD) with high homology were analyzed by ClustalX 2.1 software. Go to ExPASy website, use ProtParam tools to analyze the physical and chemical properties of CarD protein, use ProtScale to analyze protein hydrophobicity, use SignalP 4.1Server to predict protein signal peptide, use TMHMM Server v. 2.0 and TMPred to predict protein transmembrane helical region. SOPMA was used to predict protein secondary structure and SWISS-MODEL was used to model protein tertiary structure homology. Protein phosphorylation sites were predicted by NetPhos 3.1Server and conserved domains of CarD proteins were predicted by CDD tools in NCBI. Results the similarity of carD gene sequence of Mycobacterium was 100, and the evolutionary relationship was close. H37Rv and Mycobacterium vole originated from the same species and had high homology. The CarD molecule was highly conserved, stable, hydrophilic protein and no transmembrane region. There are four serine phosphorylation sites in the protein sequence without signal peptide. The secondary structure is 偽 -helix, and the tertiary structure homologous modeling is successful. CarD protein is a transcription regulator binding to RNA polymerase and belongs to the CarD/CdnL/TRCF family. Conclusion CarD protein is a RNA polymerase binding protein that regulates transcription and controls the growth of Mycobacterium tuberculosis. This protein sequence is conservative and stable, and it is a potential new target for tuberculosis treatment.
【作者單位】： 濰坊醫(yī)學(xué)院醫(yī)學(xué)檢驗(yàn)學(xué)系;濰坊醫(yī)學(xué)院臨床醫(yī)學(xué)院病原生物學(xué)教研室;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(No.30972639) 山東省自然科學(xué)基金項(xiàng)目(No.ZR2016HM09)
【分類號(hào)】：R378.911

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