人参LCAT基因的生物信息学分析及其转化胡萝卜  

Bioinformatics Analysis of Panax ginseng LCAT Gene and Its Gransformation of Daucus carota

作　　者：刘铁鹰 李纯 赵明珠 王康宇 孙春玉 陈静 王艳芳 张美萍 王义 Liu Tie ying;Li Chun;Zhao Mingzhu;Wang Kangyu;Sun Chunyu;Chen Jing;Wang Yanfang;Zhang Meiping;Wang Yi(Research Center for Ginseng Genetic Resources Development and Utilization,College of Life Science,Jilin Agricultural University,Changchun,130118)

机构地区：[1]吉林农业大学生命科学学院吉林省人参基因资源开发与利用工程研究中心

出　　处：《基因组学与应用生物学》2019年第10期4588-4595,共8页Genomics and Applied Biology

基　　金：国家863计划项目(2013AA102604);吉林省发改委-吉林省农产业创新专项资金项目(2016C04)共同资助

摘　　要：本研究对人参卵磷脂甾醇酰基转移酶基因(PgLCAT)进行生物信息学分析,以胡萝卜为受体进行遗传转化。根据已筛选的PgLCAT基因序列,利用ProtParam、ExPASy等生物信息学在线软件对PgLCAT核酸和蛋白质序列进行基本分析;使用R语言、BioLayout Express3D等软件对PgLCAT基因的表达模式进行分析;设计PgLCAT引物,以人参总RNA反转录得到的cDNA为模板,进行PCR扩增,并构建克隆载体与表达载体,进行胡萝卜转化研究。结果显示,PgLCAT基因长度为1438 bp,完整开放阅读框(ORF)的长度为1371 bp,编码457个氨基酸,相对分子质量为51473.00 kD,理论等电点为5.43,是无跨膜结构且不编码信号肽的亲水性蛋白,PgLCAT在人参14个组织部位的叶片中表达较高,其表达模式与人参皂苷的生物合成紧密相关,显著(p<0.01)影响人参单体皂苷Rb1和总皂苷的生物合成;成功构建表达载体,转化胡萝卜并获得了转基因阳性植株。为进一步深入研究PgLCAT基因的功能及其对人参皂苷生物合成机制的研究提供了科学依据。In this study,the bioinformatics analysis of ginseng lecithin cholesterol acyltransferase gene(PgLCAT)was carried out and Daucus carota as a receptor were transformed.According to the sequence of PgLCAT gene have been screened,and PgLCAT gene nucleic acid and protein sequences were analyzed by ProtParam,ExPASy and other bioinformatics online software,and the expression pattern of PgLCAT gene was analyzed by using R language,BioLayout Express3D and other software.Design specific primers,the cDNA obtained from the total RNA reverse transcriptase was used for PCR amplification,and the cloning vector and expression vector were constructed.Finally,the study on the transformation of carrots was carried out.The results showed that the length of the PgLCAT gene was 1438 bp,the length of the complete open reading frame(ORF)of the PgLCAT gene was 1371 bp,encoding 457 amino acids,molecular weight was 51473.00 kD,isoelectric point of 5.43,is a hydrophilic protein that has no transmembrane structure and does not encode signal peptide.Higher expression in leaves of the ginseng 14 tissues,the expression pattern is closely related to the biosynthesis of ginsenoside,and significantly(p<0.01)affect the biosynthesis of ginsenoside Rb1 and total saponins.It was successfully construction of cloning vector and expression vectorand transformed into carrots,and transgenic positive seedlings were obtained.For further study on PgLCAT gene function and study on ginsenoside biosynthesis mechanism provide scientific basis.

关 键 词：人参 卵磷脂甾醇酰基转移酶 胡萝卜 生物信息学分析 

分 类 号：R28[医药卫生—中药学]