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根据前期研究,进行了对比分析后明确了深绿木霉中7个敌敌畏耐受相关ABC转运蛋白,其中TaPdr2基因是在短时间内对敌敌畏胁迫应答最为明显的一个基因。本研究通过简并引物同源克隆,在深绿木霉T23中克隆出TaPdr2基因,并对其全长序列进行了测序;明确了其外显子序列和内含子序列;构建了TaPdr2的系统发生树,发现深绿木霉TaPdr2与木霉属其他种PDR5亚家族ABC转运蛋白有很高的同源性,通过生物信息学分析后预测TaPdr2蛋白的ATPase及部分跨膜结构域序列相对保守且具有11个跨膜结构域;通过构建TaPdr2的敲除载体pC1300qh-F,为后续研究TaPdr2蛋白对麦角甾醇和鞘脂类物质含量影响的研究奠定了基础。
According to the previous study, comparative analysis confirmed that seven Dichlorvos strains tolerate the ABC transporter protein, among which TaPdr2 gene is the most obvious one responding to dichlorvos stress in a short time. In this study, the TaPdr2 gene was cloned by degenerate primer homology cloning, and the full-length sequence was cloned from Trichoderma viride T23. The sequence of exon and intron was confirmed. The system of TaPdr2 The phylogenetic tree showed that Trichoderma viride TaPdr2 has high homology with other PDR5 subfamily ABC transporters of Trichoderma. After bioinformatics analysis, it is predicted that the ATPase and partial transmembrane domain sequences of TaPdr2 protein are relatively conserved It has 11 transmembrane domains. The construction of TaPdr2 knockout vector pC1300qh-F lays the foundation for further studies on the effects of TaPdr2 on the content of ergosterol and sphingolipids.