长春花细胞C_(20)hi生物碱超量合成及激素自养的分子机制探讨  

Molecular Mechanism of Alkaloid Over-Production and Hormone Autotrophy in Catharanthus Roseus Cell Line C_(20)hi

作　　者：岳庆喜[1] 祝冬青[1] 赵淑娟[1] 郑珍贵[1] 王峥涛[1] 胡之璧[1] 

机构地区：[1]国家中医药管理局中药资源与质量标准重点研究室上海市复方中药重点实验室上海中医药大学中药研究所,上海201203

出　　处：《药物生物技术》2010年第5期377-381,共5页Pharmaceutical Biotechnology

基　　金：国家自然科学基金资助课题(编号:30200358)

摘　　要：以长春花激素自养型细胞株C20hi及其母株C20D为材料,构建抑制性差减cDNA文库,斑点杂交筛选后对阳性克隆进行测序,共得到104个不同的差异表达基因片段。将这些序列在GenBank中进行BLAST比对分析,其中28个与已知基因具有很高的同源性。对部分基因的分析发现长春花细胞C20hi中g10h-like、CrPS(Catharanthus roseus protein S)、HyPRP(hybrid prolin-rich protein)、PNAE(polyneuridine aldehyde ester-ase)-like等基因的差异表达可能与该细胞株生物碱超量合成有关;而细胞色素P45071D和CR5基因的差异表达则可能与其激素自养有关。此外,培养基中2,4-D的缺失引起的TDC等酶活性的增强也可能是C20hi细胞生物碱超量合成及激素自养的原因之一。In order to make clear the molecular mechanism of alkaloid over-production and hormone autotrophy in Catharanthus roseus cell line C20hi,two Suppression Subtraction Hybridization（SSH）cDNA libraries were constructed using the cell line C20hi and its mother cell line C20D and screened applying dot blotting.As the result,104 different cDNA fragments were obtained and sequenced.Alignment by BLASTX method in GenBank revealed that 28 special sequences showed high homologue to some certain genes.These results suggested that G10H-like,HyPRP,CrPS,and PNAE-like genes may be involved in the alkaloid over-production while the cytochrome P450 71D and CR5 genes maybe related with the hormone autotrophy in the C20hi.

关 键 词：长春花 生物碱超量合成 激素自养 抑制性差减cDNA文库 

分 类 号：R979.1[医药卫生—药品] R284[医药卫生—药学]