Yocum G D
Red River Valley Agricultural Research Center, USDA-ARS, Biosciences Research Laboratory, 1605 Albrecht Boulevard, Fargo, ND 58105-5674, USA.
J Insect Physiol. 2003 Feb;49(2):161-9. doi: 10.1016/s0022-1910(02)00262-7.
Using suppressive subtractive hybridization, fragments of three diapause-associated transcripts (DAT-1, 2 and 3) were isolated from the Colorado potato beetle, Leptinotarsa decemlineata. Full-length clones were developed for the transcripts. DAT-1 encodes a deduced protein 286 amino acids in length with limited identity to several proteins with leucine-rich domains. DAT-2 encodes a deduced protein 229 amino acids in length with 27% identity, 40% similarity to the desiccation stress protein from Tenebrio molitor. DAT-3 encodes a deduced protein 97 amino acids in length with identity to no known protein. DAT-1 and 2 have similar expression patterns as determined by northern blot analysis. Trace levels of these two transcripts are first detected in 3-day-old diapause-programmed adults with a significant increase in expression on day 6. Expression of DAT-3 begins on day 12 in diapause-programmed adults and expression levels increase until the beetles enter diapause. Expression of DAT-1, 2 and 3 continues at least 60 days into diapause.
利用抑制性消减杂交技术,从科罗拉多马铃薯甲虫(Leptinotarsa decemlineata)中分离出了三个滞育相关转录本(DAT-1、2和3)的片段。为这些转录本构建了全长克隆。DAT-1编码一个推测的长度为286个氨基酸的蛋白质,与几种富含亮氨酸结构域的蛋白质具有有限的同源性。DAT-2编码一个推测的长度为229个氨基酸的蛋白质,与黄粉虫的干燥应激蛋白具有27%的同源性、40%的相似性。DAT-3编码一个推测的长度为97个氨基酸的蛋白质,与任何已知蛋白质均无同源性。通过Northern印迹分析确定,DAT-1和2具有相似的表达模式。在3日龄进入滞育程序的成虫中首次检测到这两种转录本的微量表达,在第6天表达显著增加。DAT-3的表达在进入滞育程序的成虫第12天开始,表达水平持续增加,直至甲虫进入滞育。DAT-1、2和3的表达在滞育至少60天内持续存在。