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目的:为了减少根结线虫对番茄的危害,研究并获得转抗线虫基因HS1pro1番茄植株。方法:在鉴定表达载体之后,采用CaCl2法制作农杆菌EHA105感受态细胞,然后用冻融法将HS1pro1基因转入农杆菌中。通过农杆菌介导法将HS1pro1基因导入无菌番茄外植体中,获得抗根结线虫转化再生植株。用卡那霉素筛选到再生植株后,提取抗性芽的基因组,利用设计好的引物进行PCR鉴定。结果与结论:目的基因已整合到番茄基因组中,获得了转HS1pro1基因番茄植株。
Objective: In order to reduce the damage of root-knot nematode to tomato, we studied and obtained the transgenic tomato HS1pro1 tomato plant. Methods: After identification of the expression vector, Agrobacterium tumefaciens EHA105 competent cells were made by CaCl2 method, then the HS1pro1 gene was transferred into Agrobacterium by freeze-thawing method. HS1pro1 gene was introduced into sterile tomato explants by Agrobacterium tumefaciens-mediated method to obtain transformed root-knot nematode transformed plants. After screening the regenerated plants with kanamycin, the genomes of the resistant buds were extracted and PCR was performed using the designed primers. RESULTS AND CONCLUSION: The target gene has been integrated into the tomato genome and the tomato plants transformed with HS1pro1 gene have been obtained.