Foc4 2个谷胱甘肽S-转移酶基因的克

发布时间：2018-04-20 09:04

  本文选题：Foc + 谷胱甘肽S转移酶　； 参考：《生物工程学报》2017年06期

【摘要】：为鉴定香蕉枯萎病菌(尖孢镰刀菌古巴专化型4号生理小种,Fusarium oxysporum f.sp.cubense race 4,Foc4)中的2个假想谷胱甘肽S转移酶(GSTs),采用RT-PCR方法克隆了这2个GSTs基因cDNA编码序列,随后分别将2个基因定名为Fogst1和Fogst2。其中,Fogst1的开放阅读框长609 bp,编码202个氨基酸残基,Fogst2的开放阅读框长693 bp,编码230个氨基酸残基。进化树分析表明:Fogst1属于GSTs超家族的sigma(σ)亚型成员,Fogst2属于GSTs超家族中目前未知的亚家族成员。为了验证Fogst1和Fogst2的表达,分别构建了Fogst1和Fogst2的原核表达重组载体pET28a-Fogst1和pET28a-Fogst2,并将pET28a-Fogst1和pET28a-Fogst2转化到大肠杆菌表达菌株BL21,经IPTG诱导后获得以可溶形式表达的重组蛋白Fogst1和Fogst2。GSTs活性分析表明,以CDNB为底物检测,2个重组蛋白均具有GSTs酶活性。分别取外源氧化胁迫处理后1、5、12、24 h菌丝样品进行相对荧光定量PCR分析,结果表明:Fogst1和Fogst2在前5 h表达量均大幅上调,表达量随后下调并恢复正常水平。这些结果均暗示Fogst1和Fogst2可能参与了Foc4抗外源氧化胁迫过程。
[Abstract]:In order to identify two hypothetical glutathione S-transferase (GSTs) from Fusarium oxysporum f.sp.cubense race 4 (Fusarium oxysporum f.sp.cubense race 4 Foc4), two GSTs gene cDNA coding sequences were cloned by RT-PCR. Then the two genes were named Fogst1 and Fogst2 respectively. The open reading frame length of Fogst1 was 609 BP, and that of Fogst2 encoding 202 amino acid residues was 693 BP, encoding 230 amino acid residues. The phylogenetic tree analysis shows that the member of the sigma (蟽) subtype of the GSTs superfamily belongs to the GSTs superfamily and belongs to the unknown member of the GSTs superfamily. In order to verify the expression of Fogst1 and Fogst2, the prokaryotic expression vectors pET28a-Fogst1 and pET28a-Fogst2 of Fogst1 and Fogst2 were constructed, and the pET28a-Fogst1 and pET28a-Fogst2 were transformed into E. coli expression strain BL21. The results of Fogst1 and Fogst2.GSTs activity analysis showed that the recombinant protein was expressed in soluble form after IPTG induction. Using CDNB as substrate, the two recombinant proteins had GSTs enzyme activity. The relative fluorescence quantitative PCR analysis was performed on the mycelium samples treated with exogenous oxidative stress for 24 h. The results showed that the expression of Fogst1 and Fogst2 increased significantly in the first 5 hours, then decreased and returned to normal level. These results suggest that Fogst1 and Fogst2 may be involved in the process of Foc4 resistance to exogenous oxidative stress.
【作者单位】： 海南大学热带生物资源教育部重点实验室;海南大学海洋学院;中国热带农业科学院环境与植物保护研究所;
【基金】：国家自然科学基金(No.31560491) 海南大学博士科研启动基金(No.kyqd5143)资助~~
【分类号】：S436.68

【相似文献】

相关期刊论文 前1条

1 张琛;徐辉;解书冬;贡雪峰;李亚男;樊美珍;;粉棒束孢谷胱甘肽过氧化物酶对氧化胁迫的响应[J];中国生物防治学报;2012年03期

，

本文编号：1777136