Matthews Peter R, Thornton Sarah, Gubler Frank, White Rosemary, Jacobsen John V
CSIRO Plant Industry, PO Box 1600, Canberra, ACT 2601, Australia.
CSIRO Plant Industry, PO Box 1600, Canberra, ACT 2601, Australia.Corresponding author; email:
Funct Plant Biol. 2002 Aug;29(9):1037-1043. doi: 10.1071/FP02011.
A green fluorescent protein (GFP) gene was cloned between the promoter and 3´ regions from a barley high isoelectric point (pI) α-amylase gene, then inserted into barley. GFP fluorescence was used to locate and quantify expression of the transgene in barley grains following hydration. Light and confocal laser microscopy revealed fluorescence in the known regions of α-amylase synthesis in the scutellar epithelium, aleurone layer and embryonic axis. Fluorescence was quantified using a simple fluorescence assay, which showed induction of the transgene to mirror the induction of α-amylase in aleurone exposed to gibberellic acid. Expression from the transgene was also shown to be inhibited by abscisic acid, in the same way as expression of endogenous α-amylase genes. Overall, the transgenic grain revealed patterns of α-amylase expression before and after germination, and showed strong potential for further studies investigating both α-amylase production and transport of gibberellin in malting grain.
将绿色荧光蛋白（GFP）基因克隆至大麦高等电点（pI）α-淀粉酶基因的启动子与3´区域之间，随后将其导入大麦。利用GFP荧光来定位和定量水合后大麦籽粒中转基因的表达。光学显微镜和共聚焦激光显微镜显示，盾片上皮、糊粉层和胚轴中已知的α-淀粉酶合成区域有荧光。使用简单的荧光测定法定量荧光，结果表明转基因的诱导情况反映了暴露于赤霉素的糊粉层中α-淀粉酶的诱导情况。还表明脱落酸抑制转基因的表达，方式与内源α-淀粉酶基因的表达相同。总体而言，转基因籽粒揭示了发芽前后α-淀粉酶的表达模式，并显示出在进一步研究麦芽籽粒中α-淀粉酶的产生和赤霉素转运方面具有强大潜力。
