草莓annfaf基因反义融合表达载体构建及转基因植株的获得

Construction of Anti-Sense Gene of annfaf and Genetic Transformation of Strawberry

【目的】建立研究草莓果实膜联蛋白基因annfaf功能体系。【方法】利用反义基因技术将从草莓成熟果实中分离克隆的annfaf基因定向克隆至植物中间表达载体pROKⅡ,构建了annfaf反义融合基因植物表达载体pRRJ4,通过根癌农杆菌EHA105将其导入草莓基因组。对获得的抗性植株进行PCR和Southern检测分析。【结果】表明该反义基因已整合到草莓基因组中。【结论】可进一步筛选annfaf基因表达缺陷型植株,用于探讨annfaf在非跃变型果实草莓成熟中的调控作用,为培育草莓新品种提供中间材料。

[Objective] In order to establish an experimental system for annfafgene function study in strawberry. [Method] Utilizing anti-sense technology, annfafgene which was cloned from strawberry fruit, was inserted directionally into pROK II, a plant intermediate expression vector, to construct an anti-sense fusion gene and a plant expression vector pRRJ4. Genetic transformation to strawberry was mediated by EHA105. Transgenic assays were performed using PCR and Southern blot. [Result] The anti-sense fusion annfaf gene had been integrated into strawberry genome. [Conclusion] An experimental system for selecting deficient transgenic plants which might benefit for further study on the mechanism of maturation of non-climacteric fruits such as strawberry was established. Also these deficient plants can be used as intermediate materials for fresh-keeping strawberry breeding.