Robert L S, Donaldson P A, Ladaique C, Altosaar I, Arnison P G, Fabijanski S F
Department of Biochemistry, University of Ottawa, Ont., Canada.
Plant Mol Biol. 1989 Oct;13(4):399-409. doi: 10.1007/BF00015552.
Antisense RNA was used to specifically inhibit the expression of a GUS gene introduced in a transgenic plant. A tobacco transformant containing a single intact copy of the GUS gene and showing relatively high constitutive levels of GUS activity (GUS +) was re-transformed with an Agrobacterium Ti-derived binary vector containing an antisense version of this reporter gene. The sense and antisense GUS genes were each under the regulation of the CaMV 35S promoter. Re-transformed plants contained 1-5 copies of the antisense construct and all showed a greater than 90% reduction in GUS activity relative to the original GUS + plant. This reduction in GUS activity correlated closely with the levels of GUS enzyme and steady state GUS mRNA observed in these plants. The relatively low levels of sense and antisense GUS transcripts found in the re-transformed plants may indicate a rapid degradation of the RNA:RNA duplex in the cell.
反义RNA被用于特异性抑制转基因植物中导入的GUS基因的表达。用一个含有该报告基因反义版本的根癌农杆菌Ti衍生双元载体对一个含有单个完整GUS基因拷贝且显示出相对较高组成型GUS活性水平（GUS+）的烟草转化体进行再次转化。有义GUS基因和反义GUS基因均受CaMV 35S启动子调控。再次转化的植物含有1至5个反义构建体拷贝，相对于原始的GUS+植物，所有再次转化的植物的GUS活性均降低了90%以上。GUS活性的这种降低与在这些植物中观察到的GUS酶水平和稳态GUS mRNA密切相关。在再次转化的植物中发现的有义GUS转录本和反义GUS转录本水平相对较低，这可能表明细胞中RNA:RNA双链体的快速降解。
