水稻柠檬酸合酶编码基因的初步研究

Molecular Cloning and Characterization of Citrate Synthase Gene in Rice (Oryza sativa)

以粳稻cDNA为模板,设计一对特异性引物,获得编码水稻柠檬酸合酶基因的cDNA序列,全长为1470bp,ORF区含475个氨基酸,推定蛋白序列与红葡萄、烟草、甜菜、拟南芥和柑橘等物种都有较高的同源性(>70%),氨基末端含一线粒体靶信号。Southern分析表明此基因在水稻基因组中具有多个拷贝数。经0.4mmol/LIPTG诱导,根据Western印迹分析获得预测的蛋白大小为59kD。

The full-length OsCS gene encoding citrate synthase was isolated from rice （Oryza sativa L. subsp, japonica）. OsCS is 1470 bp long and encodes a polypeptide with 475 amino acids. Its putative protein sequence is high identical to Daucus carota, N icotiana tabacum , Beta vulgaris subsp. , Arabidopsis thaliana , and Citrus j unos （ 〉 70 % ）. The deduced aminoterminal sequence of OsCS showes characteristics of mitochondrial targeting signal. Southern blot analysis using ORF of the OsCS as probe indicated that this gene exists in multiple copies in rice genome. The band with predicated size of 59 kDwas detected by Western blot after induced by 0.4 mmol/L IPTG.