Wei Zhao-Jun, Hong Gui-Yun, Wei Hong-Yi, Jiang Shao-Tong, Lu Cheng
Department of Biotechnology, Hefei University of Technology, Hefei, People's Republic of China.
DNA Seq. 2008 Jun;19(3):301-7. doi: 10.1080/10425170701605849.
Using rapid amplification of cDNA ends (RACE), the cDNA encoding eclosion hormone (EH) was cloned from the brain of Ostrinia furnacalis. The full Osf-EH cDNA is 986 bp and contains a 267 bp open reading frame encoding an 88 amino acid preprohormone, which including a hydrophobic 26 amino acid signal peptide and a 62 amino acid mature peptide. The mature Osf-EH shows high identity with Manduca sexta (95.2%), Helicoverpa armigera (91.9%) and Bombyx mori (85.5%), but low identify with Tribolium castaneum (63.6%), Drosophila melanogaster (56.5%) and Apis mellifera (54.8%). Using the HMMSTR Prediction Server, the 3D structure of Osf-EH was modeled. There are four beta-turns and three alpha-helixes predicted in Osf-EH, with the pattern of beta-beta-alpha-alpha-beta-beta-alpha. Northern blot analysis indicated a 1.0 kb transcript present only in the brain. The Osf-EH mRNA can not be detected in other neural tissues, such as the suboesophageal ganglion, thoracic ganglion, abdominal ganglion and other non-neural tissues, such as the midgut, fat body and epidermis. The Osf-EH mRNA content in the brain was measured using the combined method of quantitative RT-PCR and Southern blotting, which reached its highest level the day before the molt.
利用cDNA末端快速扩增技术(RACE),从亚洲玉米螟的脑中克隆出了编码羽化激素(EH)的cDNA。亚洲玉米螟羽化激素(Osf-EH)的完整cDNA为986 bp,包含一个267 bp的开放阅读框,编码一个由88个氨基酸组成的前体激素,其中包括一个由26个氨基酸组成的疏水信号肽和一个由62个氨基酸组成的成熟肽。成熟的Osf-EH与烟草天蛾(95.2%)、棉铃虫(91.9%)和家蚕(85.5%)具有高度同源性,但与赤拟谷盗(63.6%)、黑腹果蝇(56.5%)和意大利蜜蜂(54.8%)的同源性较低。利用HMMSTR预测服务器对Osf-EH的三维结构进行了建模。预测Osf-EH中有四个β-转角和三个α-螺旋,其模式为β-β-α-α-β-β-α。Northern杂交分析表明,1.0 kb的转录本仅存在于脑中。在其他神经组织如咽下神经节、胸神经节、腹神经节以及其他非神经组织如中肠、脂肪体和表皮中均未检测到Osf-EH mRNA。采用定量RT-PCR和Southern杂交相结合的方法测定了脑中Osf-EH mRNA的含量,其在蜕皮前一天达到最高水平。