【摘要】 以低温处理的耐盐水稻辽盐241植株叶片总RNA为模板,用OsMAPK4基因特异引物通过RT-PCR扩增出1500bp的片段,并将该片段克隆至pUC18上。序列分析结果表明,该克隆序列与GenBank上的OsMAPK4基因序列同源性达99.4%,氨基酸同源性达99.1%。进一步将OsMAPK4基因分别克隆至植物表达载体卡盒pBCE12和pBC29A上,构建了分别由E12启动子、rd29A启动子调控的OsMAPK4基因植物表达载体pBME12和pBM29A。通过冻融法将重组质粒导入根癌农杆菌LBA4404中,为农杆菌介导法转化植物,分析OsMAPK4基因的功能奠定了基础。更多还原

【Abstract】 A 1 500 bp DNA fragment amplified by RT-PCR use OsMAPK4 gene special primer from liaoyan241 leaf treated by low temperature was cloned into pUC18. The result of sequencing showed that the sequence of this fragment was 99.4% homologous with that of OsMAPK4 gene in GenBank and amino acid homologous is 99.1%. Further more OsMAPK4 gene was cloned into plant express vector cassette pBCE12 and pBC29A. Thus two plant express vectors, pBME12 and pBM29A were constructed, in which OsMAPK4 gene was controlled by E12 promoter and rd29A promoter respectively. Then the recombination plasmids were introduced into Agrobacterium LBA4404 by freezing-melting transformation method. This work provides a foundation for transferring OsMAPK4 gene into plant by Agrobacterium-mediated method and its function analysis.更多还原