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本研究以pAHC17、pCD29A质粒为基础,分别构建了玉米泛蛋白(Ubi)启动子调控的纤维素酶基因(BGLⅠ、CBHⅡ)与rd29A启动子调控的DREB1A基因的双价植物表达载体pBDB,pBDC。其植物选择标记基因为乙酰CoA转移酶基因(BAR),该载体适用于单子叶植物的遗传转化。并通过冻融法将重组质粒导入根癌农杆菌LBA4404中,为利用农杆菌介导法进行BGLⅠ,CBHⅡ和DREB1A基因对单子叶植物的基因工程研究奠定了基础。
In this study, based on the pAHC17 and pCD29A plasmids, the bivalent plant expression vectors pBDB and pBDC of DREB1A gene regulated by Ubi promoter (BGL Ⅰ and CBH Ⅱ) and rd29A promoter were constructed respectively. The plant selectable marker gene is the acetyl CoA transferase gene (BAR), which is suitable for genetic transformation of monocots. The recombinant plasmids were introduced into Agrobacterium tumefaciens LBA4404 by freeze-thaw method, which laid the foundation for the genetic engineering of monocotyledonous plants by using Agrobacterium-mediated transformation of BGLⅠ, CBHⅡ and DREB1A genes.