Imhof M O, Rusconi S, Lezzi M
Institute for Cell Biology, ETH Hönggerberg, Zürich, Switzerland.
Insect Biochem Mol Biol. 1993 Jan;23(1):115-24. doi: 10.1016/0965-1748(93)90089-b.
We have cloned a cDNA sequence coding for a Chironomus tentans steroid hormone receptor homologue which exhibits extensive amino acid sequence co-linearity with the ecdysteroid receptor of Drosophila melanogaster (dEcR; cell 67, 59-77). The DNA-binding domain has 95% and the hormone-binding domain 75% amino acid sequence identity with the cloned dEcR. The gene for this C. tentans protein is located on chromosome II, region 17C, as determined by in situ hybridization to polytene chromosomes of salivary glands. On Northern blots cDNA probes of the cloned gene hybridize to polyadenylated RNA of ca 4.2 kb. The expression of the cloned gene seems to be developmentally regulated and correlates to changes in ecdysteroid titer. Transfection of this C. tentans protein into D. melanogaster Schneider's line 2 cells leads to transcriptional interference with endogenous dEcR on an ecdysteroid-regulated promoter.
我们克隆了一段编码摇蚊类固醇激素受体同源物的cDNA序列，该同源物与黑腹果蝇的蜕皮甾体受体（dEcR；细胞67，59 - 77）表现出广泛的氨基酸序列共线性。其DNA结合结构域与克隆的dEcR具有95%的氨基酸序列同一性，激素结合结构域具有75%的氨基酸序列同一性。通过与唾液腺多线染色体的原位杂交确定，这种摇蚊蛋白的基因位于二号染色体的17C区域。在Northern印迹上，克隆基因的cDNA探针与约4.2 kb的聚腺苷酸化RNA杂交。克隆基因的表达似乎受发育调控，且与蜕皮甾体滴度的变化相关。将这种摇蚊蛋白转染到黑腹果蝇施耐德2号线细胞中，会导致对蜕皮甾体调节启动子上内源性dEcR的转录干扰。
