草鱼α-淀粉酶基因5'侧翼序列克隆与分析  

Cloning and analysis of 5' flanking sequence of α-amylase gene from Ctenopharyngodon idellus

作　　者：朱书礼[1,2] 李新辉[1] 杨计平[1] 李跃飞[1] 李捷[1] 帅方敏[1] 李琳[1] 

机构地区：[1]中国水产科学研究院珠江水产研究所,广东广州510380 [2]大连海洋大学,辽宁大连116023

出　　处：《南方水产科学》2014年第5期87-93,共7页South China Fisheries Science

基　　金：公益性行业(农业)科研专项经费项目(200903048-05);广西区自然科学基金重大项目(2013GXNSFEA053003);科技部社会公益项目(2005DIB3J023)

摘　　要：应用PCR和Genome Walking技术克隆获得长度为168 bp的草鱼（Ctenopharyngodon idellus）α-淀粉酶基因外显子Ⅰ序列和2 063 bp的5＇侧翼序列。将草鱼α-淀粉酶基因外显子Ⅰ序列与已知几种鱼类α-淀粉酶基因外显子Ⅰ序列比对,相似度为68%～86%。将草鱼α-淀粉酶基因5＇侧翼序列进行生物信息学分析,确定了其转录起始区域及转录起始位点（TSS）;在TSS上游30 bp处有1个TATA-box,-58 bp处有CCAAT-box;在5＇侧翼序列中还发现有GATA-1、OCT-1、GR、HNF-3、AP1和SP1等转录因子结合位点。草鱼α-淀粉酶基因5＇侧翼序列的克隆,为进一步研究不同食性鱼类α-淀粉酶基因侧翼序列的差异、鱼类α-淀粉酶基因的表达、功能及调控机理奠定基础。PCR and Genome Walking methods were applied to obtain a 168 bp a-amylase gene exon I sequence and a 2 063 bp α- amylase gene 5＇ flanking sequence from grass carp （ Ctenopharyngodon idellus）. The sequence alignment of a-amylase gene exon I between grass carp and other fishes showed that the similarity was 68% - 86%. The sequence analysis of a-amylase gene 5＇ flanking sequence of grass carp confirmed a transcription initiation region and a transcriptional start site （TSS）. There were a TATA-box at 30 bp and a CAAT-box at -58 bp. There were some transcriptional factor binding sites in the α-amylase gene 5＇ flanking sequence, such as GATA-1, OCT-1, GR, HNF-3, AP1 and SP1. The α-amylase gene 5＇ flanking sequence of grass carp was cloned successfully, which provides references for studies on differences of a-amylase gene 5＇ flanking sequence between different feeding fishes, as well as on the expression, function and regulation mechanism of α-amylase gene in fishes.

关 键 词：草鱼 Α-淀粉酶基因 GENOME WALKING 5'侧翼序 

分 类 号：Q953[生物学—动物学]