【摘要】：目的擴(kuò)增分枝桿菌噬菌體D29LysinB基因,對(duì)其編碼蛋白序列進(jìn)行生物信息學(xué)分析。方法提取分枝桿菌噬菌體D29基因組DNA,PCR擴(kuò)增LysinB基因并對(duì)擴(kuò)增產(chǎn)物測(cè)序;通過在線ProtParam、SOPMA、TMHMM、SWISS-MODEL等程序分析D29LysinB編碼蛋白的生物學(xué)特征。結(jié)果噬菌體D29LysinB基因全長(zhǎng)765bp,與預(yù)期大小基本一致;基因序列與Genbank上公布的MTB標(biāo)準(zhǔn)株同源性為100%。噬菌體D29LysinB蛋白共254個(gè)氨基酸,分子式為C1304H1993N349O358S7,理論等電點(diǎn)為5.78,平均疏水系數(shù)為-0.151,為親水性蛋白。二級(jí)結(jié)構(gòu)中α-螺旋、β-折疊、β-轉(zhuǎn)角和無規(guī)則卷曲分別占45.67%、10.63%、7.87%和35.83%。噬菌體D29LysinB蛋白是非跨膜或分泌蛋白。預(yù)測(cè)的B細(xì)胞抗原表位為11-28,38-46,50-55,69-75,120-127,132-139,161-171,173-177,210-213,234-237;Th細(xì)胞抗原表位為33-47,151-165,100-114,201-215,219-233,223-237,44-58,47-61,158-172,165-179,176-190,235-249;CTL細(xì)胞抗原表位為102-110,143-151,52-60,60-68,95-103,217-225,56-64,175-183,240-248,45-53,150-158,199-207。該蛋白與噬菌體Chy5LysinB蛋白進(jìn)化距離較近。結(jié)論生物學(xué)信息學(xué)方法分析噬菌體D29LysinB蛋白為親水性蛋白,含有B、Th及CTL細(xì)胞其抗原表位,為研究該蛋白在分枝桿菌中的功能奠定了基礎(chǔ)。
[Abstract]:Objective to amplify the bacteriophage D29LysinB gene of Mycobacterium and analyze its coding protein sequence by bioinformatics. Methods LysinB gene was amplified by genomic DNA,PCR from Mycobacterium bacteriophage D29 and sequenced, and the biological characteristics of D29LysinB encoded protein were analyzed by on-line ProtParam,SOPMA,TMHMM,SWISS-MODEL and other programs. Results the full length of the phage D29LysinB gene was 765 BP, which was consistent with the expected size, and the homology of the gene sequence with the MTB standard strain published on Genbank was 100%. The phage D29LysinB protein contained 254amino acids, the molecular formula was C1304H1993N349O358S7, the theoretical isoelectric point was 5.78, the average hydrophobic coefficient was-0.151, and it was a hydrophilic protein. In the secondary structure, 偽-helix, 尾-fold, 尾-turn and irregular crimping accounted for 45.67%, 10.63%, 7.87% and 35.83%, respectively. Phage D29LysinB protein is a non-transmembrane or secretory protein. B cell epitopes were predicted to be 11? 28, 38? 46, 50? 55, 69? 75120? 127132? 139161? 171173? 177210? 213234? 237?!? The antigen epitopes of CTL cells were 102? 110143? 151, 52? 60,60? 68, 95? 103217? 225, 56? 64175? 183240? 248, 45? 53150? 158199? 207? The evolution of this protein is close to that of phage Chy5LysinB protein. Conclusion Phage D29LysinB protein is a hydrophilic protein, which contains B, Th and CTL cell epitopes, which lays a foundation for studying the function of this protein in Mycobacterium. [WT5 "HZ] conclusion [WT5" HZ] [WT5 "BZ] [WT5" BZ]
【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;
【基金】：國(guó)家自然科學(xué)基金項(xiàng)目(No.81470001,No.81100006)
【分類號(hào)】：R378;Q811.4

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