Functional identification of the transcription start site and the core promoter of the juvenile hormone esterase gene in Trichoplusia ni.
Functional identification of the transcription start site and the core promoter of the juvenile hormone esterase gene in Trichoplusia ni.
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粉纹夜蛾保幼激素酯酶基因转录起始位点和核心启动子的功能鉴定。
DOI：
10.1006/bbrc.1995.2286
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发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Jones,G
中科院分区：
文献类型：
--
作者：
Schelling,D;Jones,G
将保幼激素（JH）酯酶基因克隆到T. ni编码负责降解JH的蛋白质。利用反转录PCR（RT-PCR）和北方分析对该基因的5′端结构进行了分析。用两种方法确定JHE基因的转录起始位点：一种是5′端cDNA末端快速扩增（RACE）方法，该方法产生的产物与第一内含子5′端编码的外显子序列完全相同;另一种是用内含子和外显子探针进行的北方分析。转录起始位点和包含核心启动子的区域也通过体外转录测定进行了功能鉴定。体外转录反应的产物，在推定的核心启动子区域的控制下，在RACE程序鉴定的相同碱基处起始，无论反应是由鳞翅目还是由双翅目核提取物驱动。这一结果是第一个功能鉴定的核心启动子区和转录起始位点的任何JH敏感基因。
The juvenile hormone (JH) esterase gene in T. ni encodes a protein that is responsible for the degradation of JH. The 5′ structural characterization of this JH-sensitive gene was accomplished using reverse transcription PCR (RT-PCR) and northern analysis. The transcriptional start site of the JHE gene was biochemically determined by two methods: a 5′ rapid amplification of cDNA ends (RACE) procedure which produced independent products with a sequence identical to the sequence of an exon encoded 5′ to the putative first intron and by northern analysis with intronic and exonic probes. Both the transcription start site and the region containing the core promoter were also functionally identified by use of an in vitro transcription assay. The product of the in vitro transcription reaction, under the control of the putative core promoter region, initiated at the same base as identified by the RACE procedure, whether the reaction was driven by lepidopteran or by dipteran nuclear extracts. This result is the first functional identification of the core promoter region and transcription start site of any JH-sensitive gene.