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海甘蓝总ＤＮＡ经Ｓａｕ３ＡⅠ部分酶切后，插入启动子探针型载体ｐＳＵＰＶ４的ＢａｍＨⅠ位点，转化大肠杆菌ＪＭ１０７，获得卡那霉素抗性重组质粒１３个，依次命名为ｐＲＰ１～ｐＲＰ１３。电泳分析表明它们均含有插入片段，大小为０．５～１．８Ｋｂ。各基因启动子在大肠杆菌细胞中的启动效率采用卡那霉素抗性水平测定，最高者ｐＲＰ２达１８０μｇ／ｍｌ，选择ｐＲＰ２作进一步的分析和鉴定。以ｐＲＰ２插入片段为探针，与经ＢａｍＨⅠ，ＰｓｔⅠ酶切的海甘蓝总ＤＮＡ进行Ｓｏｕｔｈｅｒｎ杂交，证明该插入片段来源于海甘蓝基因组，推测其在基因组中很可能以单考贝形式存在，斑点杂交也证明其来源。选用海甘蓝无菌苗的子叶和下胚轴为起始材料，应用根癌农杆菌（ＬＢＡ４４０４）双元载体成功地建立了一种方法简单、速度快和频率高的遗传转化体系。将ｐＲＰ２重组质粒导入海甘蓝，在附加一定量的Ａｍｐ，Ｋａｎ的相应培养基上进行筛选，２～３周就产生出卡那抗性愈伤组织和抗性小芽。
The total DNA of Chinese cabbage was partially digested with Sau3AⅠ and inserted into the BamHⅠ site of promoter-probe vector pSUPV4. The recombinant plasmid was transformed into E. coli JM107 to obtain 13 kanamycin-resistant recombinant plasmids which were named as pRP1 ~ pRP13. Electrophoresis analysis showed that they all contain insert size of 0.5 ~ 1.8Kb. The promoter efficiency of each gene promoter in E. coli cells was determined by the level of kanamycin resistance. The highest pRP2 was 180 μg / ml, and pRP2 was selected for further analysis and identification. The pRP2-inserted fragment was used as a probe to Southern hybridization with BamHI and PstI digested total kiwifruit DNA. The results showed that the inserted fragment was derived from the kiwifruit genome, Prove its source. The cotyledons and hypocotyls of aseptic seedlings of Brassica oleracea were used as starting materials. A simple, rapid and high frequency genetic transformation system was successfully established by using Agrobacterium tumefaciens (LBA4404) binary vector. The pRP2 recombinant plasmid was introduced into the sea cucumber and screened on the corresponding medium supplemented with a certain amount of Amp, Kan. After 2 to 3 weeks, kanan-resistant callus and resistant shoots were generated.