›› 2010, Vol. 37 ›› Issue (11): 61-65.

• 生物技术 • 上一篇    下一篇

鹅Pit1基因5′侧翼区克隆及序列分析

赵荣雪1，徐琪1，赵文明1，乔娜1，张海波1，段修军2，董飚2，陈国宏1   

  1. （1.扬州大学动物科学与技术学院，扬州 225009； 2.国家水禽基因库，泰州 225300）
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-11-20 发布日期:2010-11-20
  • 通讯作者: 陈国宏

Cloning and Sequence Analysis of the 5′Flaning Region of Pituitary Transcription Specific Factor 1 Gene in Goose

ZHAO Rong-xue1，XU Qi1，ZHAO Wen-ming1，QIAO Na1，ZHANG Hai-bo1，DUAN Xiu-jun2，
DONG Biao2，CHEN Guo-hong1   

  1. (1.College of Animal Science and Technology，Yangzhou University，Yangzhou 225009，China；2.National Native Waterfowl Gene Pool，Taizhou 225300，China)
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-11-20 Published:2010-11-20
  • Contact: CHEN Guo-hong

摘要： 为研究垂体特异性转录因子(pituitary specific transcription factor 1，Pit1)的启动子及其转录调控机制，试验设计了3对特异性引物，利用Genome Walking技术、克隆测序及序列比对分析，首次从太湖鹅基因组中克隆出长约3.0 kb的Pit1基因5′侧翼区序列，使用在线软件（预测侧翼序列元件）对所获序列进行分析，结果发现，Pit1包含2个启动子区域，这2个区域都含有TATA框、CAAT框和转录起始子，但无GC框，并存在一些转录因子结合位点，这些转录因子结合位点可能与潜在调控Pit1基因的转录及其表达机制有关。不同物种间Pit1基因5′侧翼区序列比对分析结果表明，Pit1基因的转录起始点上游-200 bp区域保守性较强，可能为启动子的核心序列，且鹅与鸡的同源性最高。上述结果表明，已成功获得了太湖鹅Pit1基因5′侧翼区序列。

关键词: 鹅; Pit1; 染色体步移; 基因克隆; 序列分析

Abstract: In order to explore the mechanism of Taihu goose pituitary specific transcription factor 1 (Pit1) gene transcription regulation by studying the promoter region of goose Pit1 gene，the 5′upstream sequence of Taihu goose Pit1 gene was attained by Genome Walking PCR, sequencing and sequences alignment analysis. A 3.0 kb upstream sequence of Pit1 gene was cloned firstly by Genome Walking from goose genomic template and sequenced. The sequence analysis showed that the cloned region contained two promoters， and had typical TATA box and CAAT box， but GC box was not found. Many potential transcription factors binding sits were identified by TESS analysis， which might be regulate transcription and expression the Pit1 gene potentially. The alignment results showed that the conservative region spanned from transcription start site to -200 bp of upstream，the sequence was strongly conservative and it was potential core promoter. Meanwhile，goose and chicken had higher conservative. The results showed that the cloned sequence may be considered as the promoter region of Taihu goose Pit1 gene.

Key words: goose; Pit1; Genome Walking; gene cloning; sequence analysis

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