Yang Ping, Tanaka Hiromasa, Kuwano Eiichi, Suzuki Koichi
Department of Agro-Bioscience, Faculty of Agriculture, Iwate University, Morioka 020-8550, Japan.
J Insect Physiol. 2008 Mar;54(3):636-43. doi: 10.1016/j.jinsphys.2008.01.001. Epub 2008 Jan 18.
A new cytochrome P450 gene, CYP4G25, was identified as a differentially expressed gene between the diapausing and post-diapausing pharate first instar larvae of the wild silkmoth Antheraea yamamai, using subtractive cDNA hybridization. The cDNA sequence of CYP4G25 has an open reading frame of 1674 nucleotides encoding 557 amino acid residues. Sequence analysis of the putative CYP4G25 protein disclosed the motif FXXGXRXCXG that is essential for heme binding in P450 cytochromes. Hybridization in situ demonstrated predominant expression of CYP4G25 in the integument of pharate first instar larvae. Northern blotting analysis showed an intensive signal after the initiation of diapause and no or weak expression throughout the periods of pre-diapause and post-diapause, including larval development. These results indicate that CYP4G25 is strongly associated with diapause in pharate first instar larvae.
利用消减cDNA杂交技术,在野蚕天蚕滞育和滞育后化蛹一龄幼虫中,鉴定出一个新的细胞色素P450基因CYP4G25,该基因存在差异表达。CYP4G25的cDNA序列有一个1674个核苷酸的开放阅读框,编码557个氨基酸残基。对推测的CYP4G25蛋白的序列分析揭示了基序FXXGXRXCXG,这是P450细胞色素中血红素结合所必需的。原位杂交显示CYP4G25在化蛹一龄幼虫的体表有主要表达。Northern印迹分析表明,滞育开始后有强烈信号,而在滞育前和滞育后包括幼虫发育阶段均无表达或表达较弱。这些结果表明,CYP4G25与化蛹一龄幼虫的滞育密切相关。