蓖麻RcFAH12基因cDNA全长克隆及生物信息学分析  

Full-length cDNA Cloning and Bioinformatics Analysis of RcFAH12 Gene in Castor

作　　者：狄建军[1,2,3,4,5,6] 张庆波 王月[7] 佘集凯 张树军[1,3,4,5] 黄凤兰[1,3,4,5] 李国瑞 陈永胜[1,3,4,5] 陈宇杰 Di Jianjun;Zhang Qingbo;Wang Yue;She Jikai;Zhang Shujun;Huang Fenglan;Li Guorui;Chen Yongsheng;Chen Yujie(College of Life Science, Inner Mongolia University for Nationalities, Tongliao, 028043;College of Agronomy, Shenyang Agricultural University, Shenyang, 110866;College of Agronomy, Inner Mongolia University for Nationalities, Tongliao, 028043;Engineering Research Center for Castor Industry of Inner Mongolia Universities, Tongliao, 028043;Key Laboratory for Castor Breeding of Inner Mongolia, Tongliao, 028043;Collaborate Innovation Cultivate Center for Castor Industry of Inner Mongolia, Tongliao, 028043;Key Laboratory of Bioinformatics at Inner Mongolia University for the Nationalities, Tongliao, 028043)

机构地区：[1]内蒙古民族大学生命科学学院,通辽028043 [2]沈阳农业大学农学院,沈阳110866 [3]内蒙古自治区高校蓖麻产业工程技术研究中心,通辽028043 [4]内蒙古自治区蓖麻育种重点实验室,通辽028043 [5]内蒙古自治区蓖麻产业协同创新培育中心,通辽028043 [6]内蒙古民族大学生物信息学重点实验室,通辽028043 [7]内蒙古民族大学农学院,通辽028043

出　　处：《分子植物育种》2018年第14期4583-4591,共9页Molecular Plant Breeding

基　　金：国家青年科学基金项目(31401418);国家自然科学基金项目(31160290;31460353);内蒙古自治区自然科学基金面上项目(2017MS0339);内蒙古自治区科技创新引导奖励资金项目(KJCX15002);内蒙古自治区"草原英才"计划项目(201511);内蒙古自治区草原英才创新团队支持项目(2017);内蒙古自治区高校蓖麻产业工程技术研究中心开放基金项目(MDK2017031;MDK2017030;MDK2016008);内蒙古自治区蓖麻育种重点实验室开放基金项目(MDK2016030;MDK2016031;MDK2017034;MDK2017035)共同资助

摘　　要：RACE技术是一项扩增基因末端序列的新技术。本研究以蓖麻籽的RNA为模板,以Gen Bank上FAH12基因序列设计特异引物,运用RT-PCR和RACE技术扩增并获得了特异片段。该片段经PCR、酶切和测序验证,证实所克隆序列为蓖麻FAH12的cDNA全长序列。生物信息学分析,此片段包含1 164 bp组成的开放读码框(ORF),编码387个氨基酸,分子量为44 426.21 Da,p I=8.95。同源性分析结果表明,它与麻疯树、陆地棉、百脉根和杏的FAH12基因的同源性均高于70%。构建了系统进化树,蓖麻和麻风树、橡胶、木薯聚类到一起,这与生物学分类相同,都为大戟科植物。成功克隆蓖麻FAH12基因c DNA全长,这为后续进一步的分子生物学研究提供了帮助。RACE technology is a new technology to amplify gene end sequences.In this study,castor bean RNA was used as a template to design specific primers according to the published FAH12 gene on Gen Bank.Specific primers were amplified by RT-PCR and RACE.The fragment was verified by PCR,restriction endonuclease and sequencing,which was confirmed that the cloned sequence was full-length cDNA of castor FAH12.Bioinformatics analysis showed that this fragment contained an open reading frame（ORF） of 1 164 bp,encoding 387 amino acids.Its molecular weight was 44 426.21 Da and p I=8.95.Homology analysis indicated that the homology of FAH12 gene with Jatropha curcas,Gossypium hirsutumLinn.Lotus corniculatus and Armeniaca vulgaris Lam.was higher than 70%,respectively.Phylogenetic trees were constructed,and castor and Jatropha curcas,%rubber and Manihot esculenta Crantz clustered together,which were the same as the taxonomic category and were all Euphorbiaceae plants.The successful cloning of full-length FAH12 cDNA of castor bean could provide a reference for further molecular biology research.

关 键 词：蓖麻 FAH12基因 RT-PCR RACE 克隆 

分 类 号：Q943.2[生物学—植物学] S565.6[农业科学—作物学]