果实特异性E8启动子的基因克隆

Using modified CTAB method, high purity and quality genomic DNA solution was acquired from leaf tissue, containing less polysaccharides, without browning and fit for further molecular operation. Two specific primers were designed which were added to restriction enzyme sites Hind Ⅲ and BamHⅠ. Grading-up PCR reaction system and a DNA fragment (1.1kb) of an E8 Promoter was amplified by polymerase chain reaction (PCR) and cloned into a pMD18-T Simple Vector. The plasmid DNA was isolated and identified by plasmid PCR and restriction enzyme analysis. After recombinant plasmid was identified, the results proved the correct target fragments inserted into the pMD18-T Simple Vector. Named the recombinant plasmid with pMD-E8.