【摘要】 一种新的基因克隆技术称为抑制消减杂交技术(SSH)用于研究生长于2种生态环境(古铜矿山和正常土壤)中鸭跖草Commelina communis的基因表达差异。分别以Cu矿山鸭跖草作为检测子,非Cu矿山鸭跖草作为驱赶子,建立了生长于Cu矿山生态环境中鸭跖草的差异表达cDNA文库,此cDNA文库代表在Cu矿山鸭跖草中特异表达的cDNA。通过反式Northern杂交筛选出3个阳性克隆进行测序。序列分析表明其中一个cDNA为CaM-like基因。进一步对此基因进行Virtual Northern分析,结果初步表明此基因在生长于古铜矿山上的鸭跖草中上调表达。更多还原

【Abstract】 A technique combing suppression polymerase chain reaction with suppression subtractive hybridization was used to clone low-abundance transcripts that showed differential expression between Commelina communis grown on mined area and grown on normal soil. One differential expressiong cDNA library was constructed using the cDNA of plant grown on soil area as driver and the cDNAs of plant grown on mined area as tester. This cDNA library indicated the differential expressing cDNA fragments in plant grown on mined area. By reverse Northern blot, 3 positive clones were obtained. Alignment of the three sequences were blasted in Genebank. Analysis of nucleic acid of one cDNA clone revealed that it encoded calmodulin. The calmodulin-like gene was up-regulated in the leaf of plant grown on mined area by virtual northern.更多还原