Palli S R, Ladd T R, Sohi S S, Cook B J, Retnakaran A
Molecular Entomology Laboratories, Natural Resources Canada, Ontario, Canada.
Insect Biochem Mol Biol. 1996 May;26(5):485-99. doi: 10.1016/0965-1748(96)00004-5.
Degenerate oligonucleotides and cDNA converted from Choristoneura fumiferana embryonic RNA were used in a polymerase chain reaction (PCR) procedure to isolate a 683 bp cDNA fragment. Comparison of the deduced amino acid sequence of this cDNA fragment showed that it was a region of an MHR3-like gene from C. fumeferana; we therefore named it Choristoneura hormone receptor 3 (CHR3). This CHR3 cDNA fragment was used as a probe to screen a C. fumiferana embryonic cDNA library. Twenty clones were isolated and two overlapping clones were sequenced. The longest open reading frame of CHR3 cDNA codes for 546 amino acids. The deduced amino acid sequence of this open reading frame contained all five regions typical of a steroid hormone nuclear receptor. The C domain showed the highest identity to Manduca hormone receptor 3 (MHR3), Drosophila hormone receptor 3 (DHR3) and Galleria hormone receptor 3 (GHR3). The A/B, D and E domains also showed significant amino acid similarity with MHR3, DHR3 and GHR3. The 683 bp CHR3 cDNA probe detected two mRNAs of 3.8 and 4.5 kb present during the ecdysteroid peaks for embryonic, larval, pupal and adult molts but were not detected during the intermolt periods. In sixth instar larvae, the 3.8 and 4.5 kb mRNA were detected in the epidermis, fat body and midgut tissues and the maximum expression was observed during the prepupal peak of ecdysteroids in the hemolymph. CHR3 mRNA was induced in 20-hydroxyecdysone treated CF-203 cells as well as in the midgut, fat body and epidermis of larvae that were fed the non-steroidal molting hormone agonist, RH-5992. In vitro transcription and translation of the CHR3 cDNA yielded a 61 kDa protein that bound to the retinoid related orphan receptor response element.
将来自云杉卷叶蛾胚胎RNA的简并寡核苷酸和cDNA用于聚合酶链反应（PCR）程序，以分离出一个683 bp的cDNA片段。对该cDNA片段推导的氨基酸序列进行比较表明，它是云杉卷叶蛾中一个类MHR3基因的区域；因此我们将其命名为云杉卷叶蛾激素受体3（CHR3）。这个CHR3 cDNA片段用作探针来筛选云杉卷叶蛾胚胎cDNA文库。分离出20个克隆，并对两个重叠克隆进行了测序。CHR3 cDNA的最长开放阅读框编码546个氨基酸。该开放阅读框推导的氨基酸序列包含类固醇激素核受体典型的所有五个区域。C结构域与烟草天蛾激素受体3（MHR3）、果蝇激素受体3（DHR3）和大蜡螟激素受体3（GHR3）具有最高的同一性。A/B、D和E结构域也与MHR3、DHR3和GHR3显示出显著的氨基酸相似性。683 bp的CHR3 cDNA探针检测到在胚胎、幼虫、蛹和成虫蜕皮的蜕皮甾酮峰值期间存在3.8和4.5 kb的两种mRNA，但在蜕皮间期未检测到。在六龄幼虫中，在表皮、脂肪体和中肠组织中检测到3.8和4.5 kb的mRNA，并且在血淋巴中蜕皮甾酮的预蛹峰值期间观察到最大表达。CHR3 mRNA在经20-羟基蜕皮酮处理的CF-203细胞以及喂食非甾体蜕皮激素激动剂RH-5992的幼虫的中肠、脂肪体和表皮中被诱导。CHR3 cDNA的体外转录和翻译产生了一种61 kDa的蛋白质，该蛋白质与类视黄醇相关孤儿受体反应元件结合。
