Structure and expressions of two circadian clock genes, period and timeless in the commercial silkmoth, Bombyx mori
Structure and expressions of two circadian clock genes, period and timeless in the commercial silkmoth, Bombyx mori
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DOI：
10.1016/j.jinsphys.2006.03.001
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发表时间：
2006-06-01
影响因子：
2.2
通讯作者：
Takeda, Makio
中科院分区：
文献类型：
--
作者：
Iwai, Sachio;Fukui, Yoshiko;Takeda, Makio
我们从家蚕中克隆了两个生物钟基因period（Bmper）和timeless（Bmtim）。序列分析表明，这两个基因在昆虫中具有高度的保守性。BmPER是由1，113个氨基酸组成的多肽，具有PAS、PAC、核定位信号（NLS）和胞质定位结构域（CLD）等功能结构域。推导的BmTIM由997个氨基酸组成，具有PER相互作用位点（PIS）以及NLS和CLD。Southern杂交分析表明Bmper和Bmtim是单拷贝基因。北方印迹分析表明，Bmper和Bmtim在头部和外周组织中均表达。我们还研究了Bmper和Bmtim表达的时间分布在头部，飞行肌肉，睾丸和触角下LD 12：12和LD 16：8的成年男性实时PCR测定。我们的数据表明，光周期差异影响Bmper和Bmtim的时间表达模式。LD 12：12处理下，头部Bmper和Bmtim mRNA的表达相对于LD 16：8处理有相位超前，而光周期对周围组织中Bmper和Bmtim mRNA的表达模式没有影响，光周期不仅影响相位关系，而且影响表达水平。在精巢和触角中，Bmtim的转录水平在LD 12：12中较低，但在LD 16：8中较高。Binper和Bmtim表达节律振幅的每日差异在头部为2倍，在检查的外周组织中为1.5-2.5倍。(c)2006爱思唯尔有限公司版权所有。
We cloned two circadian clock genes period (Bmper) and timeless (Bmtim) from the commercial silkmoth, Bombyx mori. Sequence analysis revealed a high degree of conservation among insects for both genes. BmPER predicted from the DNA sequence is a polypeptide of 1,113 amino acids with functional domains such as PAS, PAC, nuclear localization signal (NLS) and cytoplasmic localization domain (CLD). Deduced BmTIM consists of 997 amino acids with PER interaction site (PIS) as well as NLS and CLD. Southern blot analyses revealed that Bmper and Bmtim are single copy genes. Northern blot analysis demonstrated that Bmper and Bmtim are expressed both in the head and peripheral tissues. We also examined temporal profiles of Bmper and Bmtim expressions in the head, flight muscle, testis and antenna of adult males under LD 12:12 and LD 16:8 by Real-Time PCR assays. Our data show that photoperiod differentially affects the temporal expression patterns of Bmper and Bmtim. The mRNA expression of Bmper and Bmtim in the head had a phase lead under LD 12:12 compared to that under LD 16:8, whereas photoperiod did not affect expression patterns in peripheral tissues relative to light-on. Photoperiod affected not only the phase relationship but also the expression level. In the testis and antenna, the level of transcription of Bmtim was low in LD 12:12 but high in LD 16:8. The daily differences in amplitudes of the Binper and Bmtim expression rhythms were 2-fold in the head and 1.5-2.5 folds in the peripheral tissues examined. (c) 2006 Elsevier Ltd. All rights reserved.