利用转基因技术初步鉴定香蕉RNA解旋酶基因功能

本研究利用前期抑制差减杂交技术筛选获得的香蕉果实上调表达的基因片段,经NCBI比对,表明该基因片段属于香蕉ATP依赖的RNA解旋酶(Helicase)基因家族成员,含有完整的RNA解旋酶超家族C-末端结构域HELICc结构。经Southern杂交证实,此RNA解旋酶基因在香蕉基因组中只有一个拷贝。构建RNA解旋酶基因的反义基因植物表达载体PBIPC86,转化番茄获得6株转基因番茄植株。转基因番茄植株的叶、花和生长形态均有变异,部分转基因番茄生长势弱。该研究结果对鉴定香蕉RNA解旋酶基因功能奠定了初步基础。

Preliminary Appraisal of Banana RNA Helicase Gene Function by Transgenic Technology

In this study, an up-regulated partial gene of ATP-dependent RNA helicase from banana fruit was acquired by suppression subtractive hybridization method. Through NCBI-Blastx, it was found that the gene contained a complete RNA helicase superfamily C-terminal domain of HELICc structure. A genomic Southern Blot analysis showed that there was only one copy of the gene in the banana genome. We constructed an antisense gene plant expression vector containing an intact HELICc domain. Then we transformed this vector into tomato by using agrobacterium mediated transformation. In the transgenic tomato plants, variations happened in leaves, flowers and the growthform. The results of the study laid preliminary foundation for identifying function of banana RNA helicase gene.