小麦β-酮脂酰CoA合成酶基因KCS的克隆与酵母表达

发布时间：2018-06-03 19:51

本文选题：普通小麦 + 超长链脂肪酸　；参考：《麦类作物学报》2016年09期

【摘要】：超长链脂肪酸是生物体内众多重要物质的合成底物。KCS基因编码β-酮脂酰CoA合成酶,该酶具有底物特异性,参与超长链脂肪酸延伸的缩合反应,是超长链脂肪酸合成的限速步骤。为了探究小麦KCS基因在超长链脂肪酸合成中的功能,采用同源克隆的方法从小麦(Triticum aestivum L.)中克隆出KCS基因后,利用生物信息学对其编码序列进行分析,并在酿酒酵母(Saccharomyces cerevisiae)中对其进行真核表达。结果表明,小麦TaKCS6基因的开放阅读框为1 287bp,编码428个氨基酸残基。结构域预测结果显示,TaKCS6蛋白含有III型聚酮合酶脂肪酸延伸酶和C末端3-酰基ACP合酶III结构域,属于KCSs蛋白家族。序列比对分析结果显示,TaKCS6氨基酸序列与拟南芥及其他植物的KCS6氨基酸序列在两个功能结构域上和活性位点保守。酵母表达结果显示,TaKCS6基因编码的蛋白参与C24以上超长链脂肪酸的延伸。
[Abstract]:Ultra-long chain fatty acids (VLFAs) are the synthesis substrates of many important substances in organism. KCS gene encodes 尾 -ketolidyl CoA synthase. This enzyme is substrate-specific and participates in the condensation reaction of extended ultra-long chain fatty acids, which is the rate-limiting step for the synthesis of ultra long chain fatty acids. In order to investigate the function of wheat KCS gene in the synthesis of ultra-long chain fatty acids, a homologous cloning method was used to study the effect of KCS gene on the synthesis of ultralong chain fatty acids from Triticum aestivum L. The KCS gene was cloned and its coding sequence was analyzed by bioinformatics and expressed in Saccharomyces cerevisiae. The results showed that the open reading frame of wheat TaKCS6 gene was 1287 BP, encoding 428 amino acid residues. The results of domain prediction showed that the TaKCS6 protein contained III polyketosynthase fatty acid extender and C-terminal 3-acyl ACP synthase III domain, which belonged to the KCSs protein family. Sequence alignment analysis showed that the amino acid sequences of TaKCS6 and Arabidopsis thaliana and other plants were conserved on two functional domains and active sites. The results of yeast expression showed that the protein encoded by TaKCS6 gene was involved in the extension of ultra long chain fatty acids above C24.
【作者单位】：西北农林科技大学农学院/旱区作物逆境生物学国家重点实验室;
【基金】：国家自然科学基金项目(31471568) 陕西省重点科技创新团队项目(2014KCT-25) 西北农林科技大学唐仲英育种项目;西北农林科技大学基本科研业务费一般项目(QN2013006)
【分类号】：S512.1

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