文献类型：期刊文献

中文题名：当归甘油醛-3-磷酸脱氢酶基因的克隆及组织稳定性分析

英文题名：Gene cloning and stability analysis of glyceraldehyde-3-phosphate dehydrogenase in Angelica sinensis

作者：杨雪[1];雒军[2];杨彩霞[1];王振恒[1];夏琦[2];王引权[1]

第一作者：杨雪

机构：[1]甘肃中医药大学药学院,甘肃兰州730000;[2]甘肃中医药大学科研实验中心,甘肃兰州730000

第一机构：甘肃中医药大学药学院（西北中藏药协同创新中心办公室）

年份：2018

卷号：52

期号：6

起止页码：943

中文期刊名：河南农业大学学报

外文期刊名：Journal of Henan Agricultural University

收录：CSTPCD;;北大核心:【北大核心2017】；CSCD:【CSCD_E2017_2018】；

基金：国家重点研发计划专项"中药材生态种植技术研究及应用"项目(2017YFC1700705);国家自然科学基金项目(81660625);甘肃省高校协同创新科技团队支持计划(2016C-05FF09)

语种：中文

中文关键词：当归;GAPDH基因;克隆;序列分析;实时荧光定量PCR

外文关键词：Angelica sinensis;GAPDH gene; gene cloning;sequence analysis;qRT-PCR

摘要：通过同源比对查找GAPDH基因的保守区,运用逆转录PCR、同源克隆及cDNA末端快速克隆(rapid-amplification of cDNA ends,RACE)等技术从当归中克隆GAPDH基因的全长cDNA序列,利用生物信息学分析工具对该基因编码蛋白的基本性质、结构特点及生物学功能进行初步预测,采用实时荧光定量PCR(qRT-PCR)分析当归GAPDH基因的表达稳定性。结果表明,克隆得到当归GAPDH基因cDNA全长序列为1 345个核苷酸,开放阅读框(open reading frame,ORF)为1 005 bp,3’UTR和5’UTR长度分别为268 bp和72 bp,共编码334个氨基酸,该基因命名为AsGAPDH。AsGAPDH基因编码蛋白的预测相对分子质量为36.3 kD,理论等电点为7.06,为亲水性非分泌型蛋白,与伞形科植物白芹的亲缘关系最近。AsGAPDH基因在当归根、叶柄及叶中的表达量相近且稳定性好,扩增特异性强。
The conserved regions of GAPDH gene selected by homologous comparison was used to clone cDNA of GAPDH of Angelica sinensis through RT-PCR,homologous cloning,and rapid-amplification of cDNA ends(RACE).The expression levels of GAPDH gene were analyzed by qRT-PCR.The basic properties,features of structure,and biological functionality of encoded protein were analyzed using bioinformatics tools.The results showed that the full-length cDNA of GAPDH gene was1345bp in length,268bp3’UTR and72bp5’UTR,and a complete ORF of1005bp,which encoded334amino acids,and was designated as AsGAPDH.The predicted molecular weight of AsGAPDH protein was36.3kDa with a theoretical isoelectric point7.06showing hydrophilic non-secretory protein characteristics.AsGAPDH had the closest relationship with GAPDH of Apium graveolens.The expression levels of AsGAPDH gene in roots,petioles and leaves of Angelica sinensis were consistent,with a good stability and strong specificity.