一个NBS-LRR蛋白质基因转化水稻及其超量表达  

Transformation of a NBS-LRR Protein Gene and Its Overexpression in Rice

作　　者：郭立佳[1] 毛倩[1] 汪卫[1] 李敏[1] 彭西英[1] 陈亮[1] 

机构地区：[1]厦门大学生命科学学院细胞生物学与肿瘤细胞工程教育部重点实验室,福建厦门361005

出　　处：《厦门大学学报（自然科学版）》2010年第4期552-557,共6页Journal of Xiamen University：Natural Science

基　　金：科技部863专题(2007AA10Z132;2008ZX08001-001;2009ZX08009-045B);教育部科学技术研究重点项目(01102)

摘　　要：水稻Osxoc1334是一个编码NBS-LRR类蛋白质的基因,通过构建该基因的超量表达载体,并在农杆菌EHA105介导下转化中花11号水稻,最终获得6株再生水稻植株.对抗性愈伤组织和再生植株用GUS组织化学染色检测,结果抗性愈伤组织和6株再生水稻植株均可染上蓝色,而野生型植株不变色,用潮霉素磷酸转移酶基因的特异引物进行PCR鉴定,再生植株均可扩增到目标条带而野生型植株则不能,表明Osxoc1334基因已随T-DNA整合到再生水稻植株基因组中.采用半定量和荧光定量PCR分析转基因水稻的Osxoc1334基因表达,结果其中3株转基因植株的Osxoc1334基因平均相对表达量明显增强,其中1株为野生型植株的23.5倍.这为进一步鉴定该基因功能奠定了基础.Osxoc1334 was a NBS-LRR protein gene. Its overexpression vector was constructed and transformed rice variety Zhong-huall（Oryza sativa ssp. japonica） through Agrobacterium turnefaciens EHA105 mediated system, and thus six regenerated plantlets were obtained. The regenerated resistant embryogenic calli and plants were tested by GUS histochemical staining. As a result, all the regenerated calli and plants turned into blue but non-regenerated calli and wild type plants. These regenerated plantlets were also identified by PCR using hygromycin phosphotransferase gene specific oligonucleotide primers. The anticipated DNA fragments were amplified from all regenerated plants but wild type plant in the end. These data suggested that Oszoc1334 gene was introduced into the genome of the regenerated plants with T-DNA. The expressional levels of Osscoc1334 gene in these To plants were analyzed by semiquantitative RT-PCR and real time PCR. The expressional levels of Osxoc1334 gene in three To plants were enhanced intensively. The average relative expressional level of Osxoc1334 gene in one To plant reached about 23.5 times as compared to wild type plants.

关 键 词：抗病基因 细菌性条斑病菌 转基因水稻 基因表达 荧光定量PCR 

分 类 号：Q356.1[生物学—遗传学]