弗氏链霉菌tylF基因的克隆及其生物信息学分析

Cloning and Bioinformatics Analysis of tylF Gene in Streptomyces fradiae

[目的]研究弗氏链霉菌tylF基因的克隆及其生物信息学分析。[方法]利用RT-PCR技术、巢式PCR技术和RACE技术从弗氏链霉菌B-62169菌株中克隆获得tyl F基因的全长c DNA序列,并对其生物信息学进行分析。[结果]经Vector NTI 11.0软件拼接获得tyl F基因全长c DNA序列长度为1 245 bp,并带有19 bp长的Poly(A)尾巴,包含927 bp的开放读码框(ORF),编码一个含309个氨基酸残基的蛋白质。生物信息学分析结果表明,tyl F基因编码的酶是大菌素-O-甲基转移酶,参与分子功能中甲基转移酶活性和生物学途径中甲基化过程。对tyl F基因全长c DNA序列进行蛋白质结构域分析,证实该基因编码Tyl F蛋白,并具有223个氨基酸蛋白结构域。[结论]该研究为阐明泰乐菌素生物合成过程中甲基化反应机理,更进一步研究大环内酯类抗生素生物合成代谢途径提供生物信息学数据支持。

[Objective] To research the cloning and bioinformatics analysis of tyl F gene in Streptomyces fradiae.[Method] The partial c DNA of tyl F was cloned from Streptomyces fradiae B-62169 through RT-PCR technique by specific primers designed on the basis of tyl F gene in GenBank website.And its bioinformatics analysis was carried out.[Result]A 1245 bp full length c DNA sequence of tyl F was obtained using nested PCR and RACE technique.The cloned tyl F c DNA contained an open reading frame（ORF） of 927 bp and was predicted to encode a deduced protein with 309 amino acid residues.Bioinformatics analysis results showed that the gene encoding enzymes was macrocin-O-methyl transferase,which participated in the methylation process of methyltransferase activity and biological pathways in molecular function.Fulllength c DNA sequence of the tyl F structural gene for protein domain analysis confirmed that the gene encoding Tyl F protein having 223 amino acids was completed and the protein domains.[Conclusion] This research provides data support for the methylation reaction mechanism of tylosin biosynthetic process,and the further researches on biosynthesis and metabolic pathways of macrolides antibiotics.