采用来源于产碱杆菌(Alcaligenes eutrophus)的PHB合成酶基因phbA,经PCR扩增后,将验证正确的phbA插入到烟草质体表达载体pBio3-GFP中,取代载体中的gfp,形成prrn-phbA-aadA-TpsbA-ter表达盒,得到质体表达载体pCTHBA,通过基因枪介导,用包裹有质粒pCTHBA的金粉子弹轰击拟南芥无菌苗叶片,经壮观霉素筛选后获得拟南芥抗性植株12株;PCR验证初步表明,phbA已整合进拟南芥的质体基因组中;对转基因拟南芥植株的花器官表型和花粉显微观察表明,phbA基因在拟南芥中得到表达,显现出雄性不育的性状.%The most effective method in hybrid breeding is that many male sterility lines have been exploited for the production of F1 hybrid seeds.Based on the previously reported sequences of phbA gene, cloned from (Alcaligenes eutrophus) coding β-ketothiolase, we amplified phbA gene from plastid pBHR68 by PCR.After sequencing, digesting and ligasing, we substituted phbA gene for gfp gene in plastid vector pBio3-GFP,thus constructed plastid transformation vector pCTHBA that carries the expression cassette of prrn-phbA-aadA-TpsbA-ter.Then the vector was transformed into plastid of Arabidopsis thaliana using PDS-1000/He biolistic particle delivering system.We totally obtained 12 transgenic plants.The further PCR analysis fundamentally indicated that the phbA gene was intergrated into the plastid genome of A.thaliana.All 12 transgenic plants performed the male-sterile phenotype with 11 plants were lacking stamens and 1 plant failed to produce viable pollen by phenotype examination and microscopy scanning.
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