油桐种子FADX基因的克隆和序列分析

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。

Study on the Cloning and Sequence Analysis of FADX Gene from Vernicia fordii

Objective] The research aimed to clone full-length cDNA of FADX of Vernicia fordii 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 Vernicia fordii 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-return.Conclusion] This protein had the property of fattic acid dehydrogenase.