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当前位置: 首页 > 文章 > 文心兰OnAP1-like基因表达载体构建和遗传转化体系初探 河南农业科学 2014,43 (5) 124-129
Position: Home > Articles > Construction of Expression Vector of OnAP1-like Gene and Exploration of Genetic Transformation System of Oncidium Journal of Henan Agricultural Sciences 2014,43 (5) 124-129

文心兰OnAP1-like基因表达载体构建和遗传转化体系初探

作  者：
武振江;刘佳;崔波;叶永忠
单  位：
河南农业大学生命科学学院
关键词：
文心兰;载体构建;OnAP1基因;遗传转化体系
摘  要：
根据NCBI网站上文心兰OnAP1-like基因序列设计引物,用RT-PCR法从南茜文心兰花葶中扩增OnAP1-like基因,对扩增产物进行测序。结果表明,获得的南茜文心兰OnAP1-like基因为690bp,与报道序列完全一致。将OnAP1-like基因插入pRI101-ON载体中,经PCR、双酶切及测序鉴定,证实重组表达质粒中含有目的片段,表明成功构建了高效植物表达载体pRI101-OnAP1。对OnAP1-like基因的结构域和所表达蛋白质的亲水性进行了分析,并预测了该基因所表达蛋白的三维结构。结果显示,该基因包含MADS-box和K-box保守域,属于MADS-box基因家族,所表达蛋白为亲水性蛋白。确定了一种文心兰的遗传转化条件,得到筛选抗生素G418的最适质量浓度为40mg/L,为进一步研究文心兰中AP1基因的功能奠定基础。
译  名：
Construction of Expression Vector of OnAP1-like Gene and Exploration of Genetic Transformation System of Oncidium
作  者：
WU Zhen-jiang;LIU Jia;CUI Bo;YE Yong-zhong;College of Life Science,Henan Agricultural University;Institute of Biotechnology,Zhengzhou Normal University;
关键词：
Oncidium;;vector construction;;OnAP1 gene;;genetic transformation system
摘  要：
The OnAP1-like gene was amplified from the scape of Oncidium ‘Gower Ramsey'with primers designed according to the OnAP1-like gene sequence of Oncidiumfrom NCBI website by RT-PCR method.The PCR product was 690bp which was in fully accord with the sequence published.The OnAP1-like gene was inserted into pRI101-ON vector.The recombinant plasmid pRI101-OnAP1identified by PCR,double digestion and sequencing was constructed successfully. The multi-domain of the OnAP1-like gene and the hydrophilia of the protein encoded by the OnAP1-like gene were analyzed.The 3Dstructure of the protein encoded by the OnAP1-like gene was predicted.The results showed that this protein contained the MADS-box and K-box domain, belonging to MADS-box superfamily.The protein encoded by the OnAP1-like gene was a hydrophilic protein.The transformation system of Oncidium ‘Gower Ramsey'mediated by Agrobacterium was established,and the optimum concentration of the antibiotic G418was 40mg/ L for screening transformants.The results provide theoretical foundation for further research of the function of AP1 gene in Oncidium ‘Gower Ramsey'.

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