Expression, purification and DNA-binding activity of tilapia muscle-specific transcription factor, MyoD, produced in Escherichia coli

doi:10.1016/S1096-4959(02)00036-2

淡江大學機構典藏 > 理學院 > 生命科學研究所 > 期刊論文 > Item 987654321/61853

請使用永久網址來引用或連結此文件: https://tkuir.lib.tku.edu.tw/dspace/handle/987654321/61853

題名:	Expression, purification and DNA-binding activity of tilapia muscle-specific transcription factor, MyoD, produced in Escherichia coli
作者:	Chen, Yau-Hung;Liang, Chin-Tien;Tsai, Huai-Jen
貢獻者:	淡江大學生命科學研究所
關鍵詞:	cDNA;Fish; MyoD;Ni2+-NTA column;pET expression system;Phylogenic tree;Plaque hybridization;Tilapia
日期:	2002-04
上傳時間:	2013-06-13 11:24:03 (UTC+8)
出版者:	Philadelphia: Elsevier Inc.
摘要:	MyoD is one of several helix-loop-helix proteins regulating muscle-specific gene expression. Using a reverse transcription-polymerase chain reaction, 5′-rapid cDNA end amplification, and plaque hybridization, MyoD cDNA was cloned from the mRNA of tilapia dorsal skeletal muscle. The 1015 bp MyoD cDNA product contained an 846 bp open reading frame with flanking regions of 115 and 64 bp at the 5′- and 3′-ends, respectively. Results showed that the tilapia MyoD sequence, which includes one polypeptide of 281 amino acids, shared sequence identities of 64.3, 64.1, 62.6 and 62.4% with those of zebrafish, carp, and two rainbow trout, respectively. Results from a molecular phylogenic tree assay showed that the tilapia MyoD was more closely related to those of other fishes than of higher vertebrates. Using Escherichia coli, a pET expression system, and an Ni2+-NTA column, we purified ∼35 kDa recombinant tilapia MyoD. Results from an electrophoretic mobility shift assay demonstrated that the purified E. coli-produced tilapia MyoD was capable of binding to the DNA fragment sequence CA(C/T)(C/A)TG. © 2002 Elsevier Science Inc. All rights reserved.
關聯:	Comparative Biochemistry and Physiology Part B 131(4), pp.795-805
DOI:	10.1016/S1096-4959(02)00036-2
顯示於類別:	[生命科學研究所] 期刊論文

文件中的檔案:

檔案	描述	大小	格式	瀏覽次數
1096-4959_131_p795-805.pdf		777Kb	Adobe PDF	193	檢視/開啟
index.html		0Kb	HTML	60	檢視/開啟

在機構典藏中所有的資料項目都受到原著作權保護.

TAIR相關文章

資料載入中.....