药用猪苓菌核9种主要协助转运蛋白超家族基因的克隆与表达分析

发布时间：2018-11-26 21:40

【摘要】：目的克隆分离药用真菌猪苓菌核的主要协助转运蛋白超家族基因并进行生物信息学分析。方法采用RT-PCR技术从中国野生猪苓菌核(Polyporus umbellatus)中克隆得到9个主要协助转运蛋白超家族(major facilitator superfamily,MFS)基因,利用生物信息学方法分析这9个基因的特点,并利用qPCR验证这9个基因在不同猪苓菌核部分的表达情况。结果该9个基因cDNA包含的完整开放阅读框在1 321~1 860 bp之间,编码的氨基酸的数量在441~620之间,相对分子质量在(48.45~64.79)×10~3之间,理论等电点在6.59~9.56之间。氨基酸序列分析表明,这9个基因编码的主要协助转运蛋白的跨膜区在11~14之间。氨基酸序列多重比对及系统发育树结果显示,Comp34750、Comp34832、Comp29252、Comp42895、Comp32579和Comp27555与MFS general substrate transporter同源性比较高,Comp28872和Comp26306与MFS单糖转运蛋白(MFS monosaccharide transporter)同源性比较高,Comp33117与MFS糖转运蛋白(MFS sugar transporter)同源性比较高。实时荧光定量RT-PCR分析表明,这9个基因在不同的菌核部分都有表达,除基因Comp34382和Comp32579在共生部位显著下调外,其他7个基因都是表达显著上调。结论提示这些基因可能以不同方式参与了猪苓的防御反应及吸收外界营养元素的过程。
[Abstract]:Objective to clone and isolate the major helper transporter superfamily genes of medicinal fungus Polyporus umbellata sclerotia and analyze their bioinformatics. Methods RT-PCR technique was used to clone 9 major helper transporter superfamily (major facilitator superfamily,MFS) genes from Chinese wild Polyporus umbellata sclerotia (Polyporus umbellatus). The characteristics of the 9 genes were analyzed by bioinformatics. The expression of the nine genes in different Sclerotinia umbellata was verified by qPCR. Results the complete open reading frame contained by the 9 genes cDNA was between 1 321 and 1 860 bp, the number of amino acids encoded was between 441t620, the relative molecular weight was (48.45 ~ 64.79) 脳 10 ~ (-3), and the theoretical isoelectric point was between 6.59 and 9.56. The amino acid sequence analysis showed that the transmembrane region of the major helper transporter of these 9 genes was between 11 and 14. The results of multiple amino acid sequence alignment and phylogenetic tree showed that Comp34750,Comp34832,Comp29252,Comp42895,Comp32579 and Comp27555 had high homology with MFS general substrate transporter, and Comp28872 and Comp26306 had high homology with MFS monosaccharide transporter (MFS monosaccharide transporter). Comp33117 and MFS sugar transporter (MFS sugar transporter) have high homology. Real-time fluorescence quantitative RT-PCR analysis showed that all the 9 genes were expressed in different sclerotia. Except for the down-regulation of Comp34382 and Comp32579 in symbiotic site, the other 7 genes were up-regulated. Conclusion these genes may be involved in the defense response of Polyporus umbellata and the absorption of external nutrients in different ways.
【作者单位】：中国医学科学院北京协和医学院药用植物研究所中草药物质基础与资源利用教育部重点实验室;江苏理工学院电气信息工程学院生物信息与医药工程研究所;
【基金】：国家重点基础研究发展计划资助项目(2014CB13830X) 2014年度山西省煤基重点科技攻关项目资助项目(FT2014-03) 河北山区特色中药材种质资源评价与仿野生栽培技术研究与示范资助项目(16232503D)
【分类号】：S567.3

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