中文摘要：

以甜瓜品种‘MR-1’为试材,采用PCR技术,扩增出了At2基因并在两端添加BamHⅠ和SacⅠ酶切位点。结果表明：在NCBI上进行BLAST显示其氨基酸序列与GeneBank中已公布的At2基因的同源性为99%;经DNAMAN软件分析,核苷酸序列同源性为99.43%。将其连接在中间表达载体G-pPTN133上,得到替换了GUS基因的中间载体A-pPTN133。A-pPTN133再经ScaⅠ酶切后,插入到经ScaⅠ酶切的pPTN133~-中,构建成双T-DNA植物表达载体At2-pPTN133。经酶切和PCR鉴定证明了所构建载体的正确性。

英文摘要：

Taking melon cultivars ’MR-1’ as material,the enzymatic resistance genes At2 was amplified by PCR method and the BamHⅠenzyme site and SacⅠenzyme site was aso added.The results showed that its amino acid sequence homology was 99%compared with ＂At2＂ gene published in GeneBank after BLAST in NCBI;the nucleotide sequence homology was 99.43%after analysis by DNAMAN.The acquired gene was ligated to plasmid G-pPTN133,and therefore the GUS gene was replaced of vector A-pPTN133 was constructed The plasmid A-pPTN133 was digested by SacⅠand then ligated to the vector pPTN133~- which was aso digested by SacⅠ,so the plant expression vector At2-pPTN133 with double T-DNA was finally obtained.The results showed that enzymes digestion and PCR demonstrated the correctness of the vector.The double T-DNA vector can be transformed into melon mediated by Agrobacterium tumufaciens.