Department of Crop Sciences, University of Natural Resources and Life Sciences, Vienna, Austria.
BMC Biotechnol. 2012 Jul 2;12:37. doi: 10.1186/1472-6750-12-37.
The production of transgenic plants, either for the overproduction of the protein of interest, for promoter: reporter lines, or for the downregulation of genes is an important prerequisite in modern plant research but is also very time-consuming.
We have produced additions to the pPZP family of vectors. Vector pPZP500 (derived from pPZP200) is devoid of NotI sites and vector pPZP600 (derived from pPZP500) contains a bacterial kanamycin resistance gene. Vector pMAA-Red contains a Pdf2.1: DsRed marker and a CaMV:: GUS cassette within the T-DNA and is useful for the production of promoter: GUS lines and overexpression lines. The Pdf2.1 promoter is expressed in seeds and syncytia induced by the beet cyst nematode Heterodera schachti in Arabidopsis roots. Transgenic seeds show red fluorescence which can be used for selection and the fluorescence level is indicative of the expression level of the transgene. The advantage is that plants can be grown on soil and that expression of the marker can be directly screened at the seed stage which saves time and resources. Due to the expression of the Pdf2.1: DsRed marker in syncytia, the vector is especially useful for the expression of a gene of interest in syncytia.
The vector pMAA-Red allows for fast and easy production of transgenic Arabidopsis plants with a strong expression level of the gene of interest.
生产转基因植物,无论是为了过量生产感兴趣的蛋白质,为启动子:报告基因系,还是为了下调基因,都是现代植物研究的重要前提,但也非常耗时。
我们已经生产了 pPZP 家族载体的添加物。载体 pPZP500(源自 pPZP200)不含 NotI 位点,载体 pPZP600(源自 pPZP500)含有细菌卡那霉素抗性基因。载体 pMAA-Red 在 T-DNA 内包含了 Pdf2.1:DsRed 标记和 CaMV::GUS 盒,可用于生产启动子:GUS 系和过表达系。Pdf2.1 启动子在种子和甜菜胞囊线虫 Heterodera schachti 在拟南芥根中诱导的合胞体中表达。转基因种子显示红色荧光,可用于选择,荧光水平表明转基因的表达水平。其优点是植物可以在土壤中生长,并且可以在种子阶段直接筛选标记的表达,从而节省时间和资源。由于 Pdf2.1:DsRed 标记在合胞体中的表达,该载体特别适用于在合胞体中表达感兴趣的基因。
载体 pMAA-Red 允许快速、轻松地生产具有强表达水平的感兴趣基因的转基因拟南芥植物。