Manteuffel R, Panitz R
Institute of Plant Genetics and Crop Plant Research, Gatersleben, Sachsen-Anhalt, Germany.
Plant Mol Biol. 1993 Sep;22(6):1129-34. doi: 10.1007/BF00028981.
We have used a highly sensitive immunological tissue print technique to study cell- and tissue-specific expression of heterologous genes in transgenic plants. Primary polyclonal antibodies, raised against legumin of faba bean (Vicia faba L.) and 12S globulin of oat (Avena sativa L.) were used to localize these proteins in transgenic tobacco seeds in a streptavidin-alkaline phosphatase assay in combination with biotinylated secondary antibodies producing a higher sensitivity (by several amplification steps) of the assay. Both storage protein genes were found to be expressed in a specific pattern. While legumin is preferentially accumulated in certain parts of the embryo, the oat legumin-type globulin is restricted to the endosperm. The applied technique is highly sensitive with a resolution power down to the single-cell level and allows rapid screening of large numbers of samples.
我们采用了一种高度灵敏的免疫组织印迹技术来研究转基因植物中异源基因的细胞和组织特异性表达。针对蚕豆（Vicia faba L.）的豆球蛋白和燕麦（Avena sativa L.）的12S球蛋白产生的初级多克隆抗体，用于在链霉亲和素 - 碱性磷酸酶测定中结合生物素化二抗，在转基因烟草种子中定位这些蛋白质，从而提高了测定的灵敏度（通过几个放大步骤）。发现这两个贮藏蛋白基因均以特定模式表达。豆球蛋白优先积累在胚的某些部位，而燕麦豆球蛋白型球蛋白则局限于胚乳。所应用的技术高度灵敏，分辨率可达单细胞水平，并允许快速筛选大量样品。
