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【目的】筛选阳春砂1-去氧-D-木酮糖-5-磷酸还原异构酶(DXR)(AvDXR)转基因烟草T1代植株,鉴定AvDXR基因在模式植物烟草中的功能。【方法】采用抗生素、聚合酶链反应(PCR)法筛选转基因烟草T1代植株;采用半定量逆转录—聚合酶链反应(RT-PCR)法分析目的基因在T1代转基因烟草中的表达情况,采用分光光度法测定酶活性和光合色素含量。【结果】经过抗生素筛选和PCR检测获得AvDXR基因稳定遗传的转基因烟草T1代;AvDXR在部分T1代植株中得到超表达;经测定超表达植株中的DXR活性和光合色素含量均有提高。【结论】AvDXR超表达提高了转基因烟草DXR活性和光合色素含量,获得的AvDXR超表达植株可作为进一步分析萜类成分的材料。
【Objective】 The purpose of this study was to screen the plants of T1 generation of transgenic plants of 1-deoxy-D-xylulose-5-phosphate reductoisomerase (DXR) (AvDXR) of Populus simonii to identify the function of AvDXR gene in model plant tobacco. 【Method】 The T1 generation transgenic tobacco plants were screened by antibiotic and polymerase chain reaction (PCR). The expression of the target gene in T1 generation transgenic tobacco was analyzed by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR) Spectrophotometric determination of enzyme activity and photosynthetic pigment content. 【Result】 T1 generation of transgenic tobacco with stable AvDXR gene was obtained through antibiotic screening and PCR detection. AvDXR was overexpressed in some T1 plants. DXR activity and photosynthetic pigment content of over-expressed plants were all increased. 【Conclusion】 AvDXR overexpression increased DXR activity and photosynthetic pigment content of transgenic tobacco, and the obtained AvDXR overexpression plants could be used as materials for further analysis of terpenoid components.