The fat body cell-free system for tissue-specific transcription of plasma protein gene of Bombyx mori.
The fat body cell-free system for tissue-specific transcription of plasma protein gene of Bombyx mori.
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用于家蚕血浆蛋白基因组织特异性转录的脂肪体无细胞系统。
DOI：
10.1093/nar/23.14.2648
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发表时间：
1995
影响因子：
14.9
通讯作者：
S. Tomino
中科院分区：
文献类型：
--
作者：
E. Mine;H. Sakurai;S. Izumi;S. Tomino
制备了家蚕幼虫脂肪体的核提取液，建立了家蚕主要血浆蛋白基因转录的体外同源体系。在较高的模板浓度下，家蚕贮藏蛋白SP1基因和腺病毒2主要晚期(Adml)基因在5龄雌幼虫脂肪体提取液中得到了忠实转录。低浓度α-Amanitin对基因转录的完全抑制表明，该反应是由RNA聚合酶II催化的。在低模板浓度(0.6 nM)下，脂肪体核提取液高效转录同源SP1基因，而Adm1基因和幼虫角质层蛋白基因仅在同一提取液中勉强转录。TATA盒序列上游缺失的SP1基因对转录影响不大，而破坏TATA序列的突变完全取消了该基因的转录。这些结果表明，SP1基因的核心启动子区域位于-44和+16之间，是脂肪体体外特异转录所必需的。
A nuclear extract was prepared for the larval fat body of the silkworm, Bombyx mori, and a homologous in vitro system was developed for the transcription of major plasma protein gene of B.mori. The gene for SP1, a storage protein of B.mori, and adenovirus 2 major late (AdML) gene were faithfully transcribed under relatively high template concentrations in the nuclear extract prepared from the fat body of female fifth instar larvae. Complete inhibition of gene transcription by a low concentration of alpha-amanitin indicated that the reaction is catalyzed by RNA polymerase II. At low template concentration (0.6 nM) the fat body nuclear extract transcribed the homologous SP1 gene with high efficiency, while AdML gene and larval cuticle protein gene were only barely transcribed in the same extract. The SP1 gene deleted upstream of the TATA box sequence showed little effect on transcription, whereas mutations that destroy TATA sequence totally abolished the gene transcription. These results suggested that the core promoter region of SP1 gene spanning between positions -44 and +16 is essential for the fat body specific transcription in vitro.