太子参GAPDH基因的克隆及序列分析  

Cloning and Sequence Analysis of GAPDH Gene from Pseudostellaria heterophylla

作　　者：龙登凯 丁铃[1] 李军[1] 周涛[1] 江维克[1] 肖承鸿[1] 

机构地区：[1]贵阳中医学院,贵州贵阳550002

出　　处：《河南农业科学》2016年第7期87-92,共6页Journal of Henan Agricultural Sciences

基　　金：国家自然科学基金项目(81460579);贵州省研究生工作站建设项目(黔教研合JYSZ字[2014]016);施秉中药材产业科技合作专项计划项目[施中药科合专项(2014)第6号]

摘　　要：为了获取可靠内参基因,采用同源克隆技术和c DNA末端快速延伸技术,首次从太子参中克隆3-磷酸甘油醛脱氢酶基因(GAPDH)的部分c DNA序列,命名为Ph GAPDH,序列长度为981 bp,包含699 bp的部分阅读框,推测其编码232个氨基酸,还含有282 bp的3'UTR。序列分析结果表明,太子参Ph GAPDH基因编码的氨基酸序列与同科植物香石竹(Dianthus caryophyllus)氨基酸序列同源性高达99%。PCR分析结果表明,Ph GAPDH基因在太子参不同种源块根、同一种源不同组织及不同生长时期的表达水平较稳定,可以作为太子参功能基因研究的内部参考基因。In order to achieve a valuable reference gene,a partial c DNA sequence of GAPDH was cloned from Pseudostellaria heterophylla by the homology cloning and RACE,which was named Ph GAPDH. The length of Ph GAPDH c DNA was 981 bp,which encoded 232 amino acids with 282 bp 3’ UTR. Sequence analysis indicated that the amino acid of Ph GAPDH showed high similarity with the GAPDH from Dianthus caryophyllus（ 99%）. The PCR analysis showed that Ph GAPDH had a consistent expression in roots from different provenances and in different tissues,growth stages. This result showed that Ph GAPDH could be used as a reliable reference gene in the function research of genes from P. heterophylla.

关 键 词：太子参 GAPDH基因 同源克隆法 CDNA末端快速扩增 表达模式 

分 类 号：Q785[生物学—分子生物学]