Livingstone Donald, Beilinson Vadim, Kalyaeva Marina, Schmidt Monica A, Herman Eliot M, Nielsen Niels C
Department of Agronomy, Purdue University, West Lafayette, IN 47907, USA.
Plant Mol Biol. 2007 Jul;64(4):397-408. doi: 10.1007/s11103-007-9163-x. Epub 2007 Apr 11.
A mutant Bowman-Birk gene was created that encoded an inactive high-sulfur product. It was used to transform soybean line Asgrow 3237. Transformants bearing the mutant gene were identified by GUS expression, PCR analysis, and Southern analysis. The amount of steady state mRNA from the mutant gene in the transformed plants showed that the gene was highly expressed, but the amount of message from the unmodified Bowman-Birk gene did not change detectably. Proteins synthesized at the direction of the mutant Bowman-Birk gene accumulated in seeds of the transformed plants, and there was a marked decrease in the ability of extracts prepared from these seeds to inhibit trypsin and chymotrypsin despite the presence of Kunitz trypsin inhibitor. The more prevalent mRNA from the mutant gene was considered to out-compete message from the native genes to decrease the amount of active Bowman-Birk inhibitor.
构建了一个编码无活性高硫产物的突变型鲍曼-伯克基因。该基因用于转化大豆品系Asgrow 3237。通过GUS表达、PCR分析和Southern分析鉴定携带突变基因的转化体。转化植株中突变基因的稳态mRNA量表明该基因高表达，但未修饰的鲍曼-伯克基因的mRNA量没有明显变化。在突变型鲍曼-伯克基因指导下合成的蛋白质在转化植株的种子中积累，尽管存在库尼茨胰蛋白酶抑制剂，但从这些种子制备的提取物抑制胰蛋白酶和胰凝乳蛋白酶的能力显著下降。来自突变基因的更普遍的mRNA被认为与来自天然基因的mRNA竞争，从而减少活性鲍曼-伯克抑制剂的量。
