草鱼C–反应蛋白基因上游调控序列的扩增与分析

Amplification and analysis of up-stream regulatory sequence of C–reactive protein gene(CRP) from grass carp(Ctenopharyngodon idellus)

C-反应蛋白（CRP）是一种急性时相血清蛋白，与鱼类的天然免疫和炎症反应有密切的关系。通过Genome Walker方法扩增草鱼CRP基因的上游序列，获得长度为1055bp的DNA片段，测序后经过PLACE和BDGP等启动子和转录因子结合位点预测软件的预测，初步鉴定草鱼CRP基因的复制起始子序列为保守的TCAGATC，G为转录起始位点。高度保守的RNA聚合酶11的结合位点TATAA-box位于-41- -35bp，CAAT-box位于-94- -91bp，在-54— -5bp处存在1个高度保守的核心启动子序列。此外，还发现与转录诱导调控有关的转录因子结合位点，包括4个E-box元件、3个GT1共有序列、2个GT1核心序列和2个I-box核心序列。

C-reactive protein （CRP） is a kind of acute phase serum protein associated with innate immunity and inflammatory reaction offish. A 1 055 bp DNA fragment from grass carp CRP gene up-stream sequence was amplified by Genome Walker. Sequence analyzing by online software PLACE and BDGP for prediction of promoter sequence and transtription factor binding sites indicated that the conserved initiate sequence of grass carp CRP gene is TCAGATC and the initiate site of transcription is G. RNA polymerase II binding site, TATA-Box, is located in -41 to -35 bp which embed in a highly conserved core promoter sequence （-54 to -5 bp）, CAAT-Box appears in -94 to -91 bp. Moreover, some transcriptional factor binding sites related to inducible regulation of transcription are found in up-stream sequence, including 4 E-box elements, 3 GT1 consensus sequences, 2 GTl-core elements, and 2 I-box-core elements.