竹黄菌veA基因全长克隆与序列分析

发布时间：2018-06-25 05:11

本文选题：竹黄菌 + veA基因　；参考：《基因组学与应用生物学》2017年09期

【摘要】：为了研究ve A基因在竹黄菌中的调节机制,利用RT-PCR(reverse transcriptase-PCR)、RACE(rapid amplification of c DNA ends)及染色体步移技术克隆获得了ve A基因的全长序列。序列分析显示:该基因序列全长为1 780 bp,其开放阅读框长度为1 104 bp,编码367个氨基酸;其编码的蛋白质分子量为40.887 5 k D,理论等电点p I为8.95,含42个负电荷氨基酸和46个正电荷氨基酸,是一种弱酸性蛋白;序列同源性分析结果显示:该基因编码的蛋白序列与偃麦草核腔菌(Pyrenophora tritici-repentis)的ve A蛋白序列(XM_001933944.1)及异旋孢腔菌(Cochliobolus heterostrophus)的velvet-like protein(JF826791.1)的同源性最高,其identity分别为78%和79%;对该基因编码的蛋白质进行二级结构预测,结果显示:该蛋白α-螺旋(alpha helix)占11.72%,延伸链(extended strand)占24.52%,β-转角(beta turn)占9.54%,无规卷曲(random coil)占54.22%;对其保守结构域分析显示,该蛋白含有一个Velvet superfamily结构域;利用同源模建方法预测其三级结构,结果显示该蛋白的三级结构模型与隔孢腔菌目中其他物种的ve A异源二聚体的相似性为46%。
[Abstract]:In order to study the regulation mechanism of VE A gene in bamboo, the full-length sequence of the gene was obtained by RT-PCR (reverse transcriptase-PCR), RACE (rapid amplification of C DNA ends) and chromosome step technique. The sequence analysis showed that the length of the gene sequence was 1780, and the length of the open reading frame was 1104 and 367 encoded. The molecular weight of the protein is 40.8875 K D, the theoretical isoelectric point P I is 8.95, with 42 negative charge amino acids and 46 positive charge amino acids, which is a weak acid protein; sequence homology analysis results show that the gene sequence of the gene and the ve A protein sequence of Pyrenophora tritici-repentis (XM_0) is the sequence of VE A protein (XM_0). 1933944.1) and the homology of velvet-like protein (JF826791.1) of Cochliobolus HETEROSTROPHUS was the highest, and its identity was 78% and 79%, respectively. The protein encoded by this gene was predicted by the two grade structure. The results showed that the protein alpha helix (alpha helix) accounted for 11.72%, the elongation chain (extended strand) accounted for 24.52%, and beta angle (BET) A turn) accounted for 9.54%, and the random curl (random coil) accounted for 54.22%. Its conservative domain analysis showed that the protein contained a Velvet superfamily domain, and the homologous model building method was used to predict the three-stage structure. The results showed that the similarity between the three structure model of the protein and the VE A heterogenous two polymer in other species of the septum was 46%.
【作者单位】：贵州省生物技术研究所;贵州省农业生物技术重点实验室;
【基金】：国家自然科学基金项目“竹黄菌侵染桂竹互作机制研究”(NSFC 31460199) 贵州省科技合作计划项目“竹黄菌产竹红菌素相关基因的功能验证”黔科合LH字(2015)7064号共同资助
【分类号】：Q78;S567.39

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