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通过mRNA差异显示技术从豆蝽Riptortus clavatus脂肪体中鉴定并克隆新型保幼激素响应基因

Identification and cDNA cloning of novel juvenile hormone responsive genes from fat body of the bean bug, Riptortus clavatus by mRNA differential display.

作者信息

Hirai M, Yuda M, Shinoda T, Chinzei Y

机构信息

Department of Medical Zoology, School of Medicine, Mie University, Tsu, Japan.

出版信息

Insect Biochem Mol Biol. 1998 Mar;28(3):181-9. doi: 10.1016/s0965-1748(97)00116-1.

Abstract

Juvenile hormone (JH) induces termination of diapause and initiation of reproductive maturation in the adult female bean bug, Riportus clavatus. Applying PCR-based differential display, we have identified four novel JH-responsive cDNAS, that is, three repressible (JR-1, 2 and 3) and one inducible (JI-1). These DNA fragments were partially sequenced and compared with sequences in the data base. JR-3 was shown to have similarity to the transferrins of other insects, which have been reported as JH-suppressed genes. JI-1 has similarity to vitellogenin of Aedes aegypti. On the other hand, JR-1-1 and 2 have no significant similarity to other known sequences. For JR-1, the full cDNA sequence was determined: it contained 913 bp, encoding 194 amino acid residues with a calculated M.W. of 21,531 Da in the mature protein. A total of six JH-responsive genes or cDNAs (four suppressible and two inducible or stimulated), including the already isolated JH-responsive cyanoprotein genes (CP-alpha and beta), have been isolated and are available for further comparative analysis of gene structure and regulatory mechanisms in the same tissue under the same hormonal conditions.

摘要

保幼激素(JH)可诱导成年雌性豆蝽(Riportus clavatus)滞育的终止和生殖成熟的启动。应用基于聚合酶链反应(PCR)的差异显示技术,我们鉴定出了四个新的JH反应性cDNA,即三个受抑制的(JR-1、2和3)和一个诱导性的(JI-1)。对这些DNA片段进行了部分测序,并与数据库中的序列进行了比较。结果显示JR-3与其他昆虫的转铁蛋白具有相似性,这些转铁蛋白已被报道为JH抑制基因。JI-1与埃及伊蚊的卵黄原蛋白具有相似性。另一方面,JR-1和2与其他已知序列没有显著的相似性。对于JR-1,我们测定了其完整的cDNA序列:它包含913个碱基对,编码194个氨基酸残基,成熟蛋白的计算分子量为21,531道尔顿。总共六个JH反应性基因或cDNA(四个受抑制的和两个诱导性或刺激性的),包括已经分离出的JH反应性蓝蛋白基因(CP-α和β),已被分离出来,可用于在相同激素条件下对同一组织中的基因结构和调控机制进行进一步的比较分析。

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