Mouillet J F, Delbecque J P, Quennedey B, Delachambre J
Institut für Zellbiologie, ETH-Hönggerberg, Zürich, Switzerland.
Eur J Biochem. 1997 Sep 15;248(3):856-63. doi: 10.1111/j.1432-1033.1997.00856.x.
Using the Drosophila EcR-B1 cDNA as a probe, we have cloned the putative ecdysteroid receptor from the mealworm Tenebrio molitor. We have isolated two cDNAs with different 5' termini that contain a complete open reading frame. These two cDNAs encode two proteins with distinct N-terminal regions corresponding to two isoforms. The coleopteran receptor is obviously related to the ecdysteroid receptor of other insects, but shares only 89% and 61% amino acid identities with the DNA-binding and ligand-binding domains of the Drosophila receptor, respectively. Its expression pattern has been examined in the epidermis during the last larval instar and pupal stage of T. molitor, in correlation with the hemolymph ecdysteroid titer. Hybridizations revealed two transcripts of 7 kb and 6.5 kb detected in most stages during metamorphosis and corresponding to the A and B1 isoforms. These two mRNAs are highly evident just before the rise of each ecdysteroid peak both in prepupae and in pupae. They show almost the same expression pattern in epidermis except for the second part of the pupal stage, during which only the A isoform is detected.
以果蝇EcR - B1 cDNA为探针,我们从黄粉虫(Tenebrio molitor)中克隆了假定的蜕皮甾体受体。我们分离出了两个5'末端不同但包含完整开放阅读框的cDNA。这两个cDNA编码两种蛋白质,它们具有不同的N端区域,对应于两种异构体。鞘翅目受体显然与其他昆虫的蜕皮甾体受体相关,但与果蝇受体的DNA结合域和配体结合域的氨基酸同一性分别仅为89%和61%。在黄粉虫最后一龄幼虫和蛹期的表皮中,我们检测了其表达模式,并与血淋巴蜕皮甾体滴度相关联。杂交显示在变态过程的大多数阶段检测到7 kb和6 .5 kb的两种转录本,分别对应于A和B1异构体。在预蛹和蛹中,这两种mRNA在每个蜕皮甾体峰上升之前都非常明显。除了蛹期的第二部分,在该阶段仅检测到A异构体,它们在表皮中的表达模式几乎相同。