樱桃ACC氧化酶基因的克隆和序列分析

Cloning and Sequencing of ACC Oxidase Genes in Cherry Fruit and Leaf

以樱桃基因组DNA为模板定向克隆其ACC氧化酶基因。对PCR扩增的目的片段用 1%琼脂糖凝胶电泳检测后 ,进行回收、连续转化 ,通过蓝白筛选挑取白色菌落提取质粒DNA进行酶切鉴定 ,确定重组子后采用双脱氧终止法测定DNA全序列。DNA测序结果显示 ,樱桃的ACC氧化酶基因序列全长为 1310bp ,由 2个外显子和 3个内含子构成 ,外显子总长为 10 0 0bp。同源性分析可知 ,樱桃与桃的ACC氧化酶基因DNA序列同源性达 97 8%。

The ACC oxidase gene was cloned from genomic DNA of prunus pseudocerasus L.Destination segments amplified by PCR were reclaimed,connected and transformed after examination by 1% agarose gel electrophoresis.By blue white screening,the plasmid DNA extracted from white colony was identified using restriction enzyme analysis and the DNA sequence was determined following recon determination.The results of DNA sequencing revealed that the ACC oxidase gene sequence of Prunus pseudocerasus L was 1 310 bp and consisted of 2 exons and 3 introns and the full length of the exons was 1 000 bp.Homogenous analysis indicated that homologue of DNA sequence of the ACC oxidase gene in Prunus pseudocerasus L and Prunus persica Batsch reached 97 8%.