水稻钙依赖型蛋白激酶OsCPK9基因启动子的克隆及在烟草中的瞬时表达分析

韦淑亚，刘小东，张莹莹，赵旭东，罗青晨，刘虎，杨广笑*，何光源*

Abstract: Calciumdependent protein kinases (CDPKs) regulate the downstream components in calcium signaling pathways and play important roles during the growth and development of plant and in response to various biotic and abiotic stresses. Based on the cDNA sequence of OsCPK9， a CDPK gene in rice， a 2 kb putative promoter sequence was located in rice genome by searching NCBI database， and was amplified by PCR using genomic DNA of rice (Oryza sativa L. cv.Nipponbare) as template and specific primers designed according to the predicted sequence. The cloned OsCPK9 promoter was 2 114 bp in length，and designated POsCPK9. PLANTCARE online software analysis indicated that POsCPK9 contains some cisacting expression elements related to stress signals， except basic elements TATAbox and CAATbox. A POsCPK9 and GUS fusion expression vector POsCPK9GUS was constructed by replacing CaMV 35S promoter of pBI121 with the POsCPK9. This vector was transiently expressed in the tobacco plant through Agrobacteriummediated transformation method. The histochemical GUS staining and activity analysis showed that GUS could be expressed in roots， stems and leaves of tobacco plant with different levels， which suggested the upstream 2 kb sequence of OsCPK9 gene has promoter function.

Link: http://www.zjnyxb.cn/CN/Y2014/V26/I2/0