摘要
[目的]克隆油桐种子FADX基因全长cDNA。对该基因作生物信息学分析,为进一步研究该基因的功能提供参考。[方法]以未成熟的油桐种子为材料,利用改良TRIzoL法提取总RNA,并根据GenBank中已经登录的α桐酸合成酶基因FADX的序列,设计特异性引物;采用RTPCR方法,克隆得到FADX基因全长cDNA,应用Antheprot软件和InterProScan对该基因进行了生物信息学分析。[结果]该序列5'端和3'端非编码区序列长度分别为13、47 bp,含有1个开放阅读框(14～1 174 bp),编码386个氨基酸,含有典型的脂肪酸脱氢酶结构域;相对分子量是44 343.0,理论等电点为8.33。二级结构预测表明,该蛋白α螺旋含量占29%,β折叠占32%,转角占6%。[结论]该蛋白属于脂肪酸脱氢酶。
[Objective] The research aimed to clone full-length cDNA of FADX of Verniciafordii seeds. The bioinformatic analysis of this gene was done, and provided reference for the further research of the function of gene. [Method] Total RNA were successfully extracted by developing Verniciafordii seeds with improved TRIzoL method. Primer was designed according to the FADX sequences recorded in Genbank. Full-length cDNA of FADX was obtained by cloning it with RT -PCR method. Bioinformatic analysis were done by using the software of Antheprot and Interproscan. [Result] The length of 5＇ side and 3＇ side unencoded area＇s sequences were 13 bp and 47 bp, contained an opening reading frame from 14 bp to 1 174 bp which encoded a polypeptide of 386 amino acids, and contained a specific fattic acid dehydrogenase structural domain. Relative molecular weight was 44 343.0, pI was 8.33, and the calculation of secondary structure showed that the protein contained 29% in α - helix, 32% in β-pleated sheet, and 6% inβ-returu.[Conclusion] This protein had the property of fattic acid dehydrogenase.
出处
《安徽农业科学》
CAS
北大核心
2008年第11期4753-4755,共3页
Journal of Anhui Agricultural Sciences
基金
浙江省重大科技项目"能源植物种质资源与高能植物品种选育及中试"(2006#2009)(2005C12003)
南京中医药大学科技发展基金项目"生物柴油原料植物油桐脂肪酸合成酶相关基因克隆和功能分析"(2006-2007)
