Zebrafish Sp1-like protein is structurally and functionally comparable to human Sp1

Cha Jang Lin, Tsun Hsien Hsiao, Yi Shao Chung, Wen Ni Chang, Trai-Ming Yeh, Bing Hung Chen, Tzu-Fun Fu

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

The transcription factor Sp1 is a regulator of TATA-less genes. It belongs to the Cys2-His2 zinc finger domain-containing family. A zebrafish cDNA encoding a peptide homologous to mammalian Sp1 was cloned and inserted into a pET43.1a vector and expressed in Escherichia coli Rosetta (DE3) cells as a Nus-His-tag fusion protein. After induction with isopropyl thiogalactoside, the protein was purified with a Ni-Sepharose column, and approximately 5-8 mg of pure protein was obtained per liter of culture. The primary sequence and the predicted partial tertiary structure of the potential recombinant zebrafish Sp1 protein are similar to those of human Sp1. The DNA affinity precipitation assay and dual-luciferase promoter activity assay further confirm the nature of the recombinant zebrafish Sp1 protein as a transcription factor. Our results show that zebrafish Sp1-like protein is structurally and functionally comparable to human Sp1.

Original languageEnglish
Pages (from-to)36-43
Number of pages8
JournalProtein Expression and Purification
Volume76
Issue number1
DOIs
Publication statusPublished - 2011 Mar 1

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Zebrafish
Zebrafish Proteins
Recombinant Proteins
Proteins
Sp1 Transcription Factor
Isopropyl Thiogalactoside
Zinc Fingers
Luciferases
Sepharose
Transcription Factors
Complementary DNA
Escherichia coli
Peptides
DNA
Genes

All Science Journal Classification (ASJC) codes

  • Biotechnology

Cite this

Lin, Cha Jang ; Hsiao, Tsun Hsien ; Chung, Yi Shao ; Chang, Wen Ni ; Yeh, Trai-Ming ; Chen, Bing Hung ; Fu, Tzu-Fun. / Zebrafish Sp1-like protein is structurally and functionally comparable to human Sp1. In: Protein Expression and Purification. 2011 ; Vol. 76, No. 1. pp. 36-43.
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abstract = "The transcription factor Sp1 is a regulator of TATA-less genes. It belongs to the Cys2-His2 zinc finger domain-containing family. A zebrafish cDNA encoding a peptide homologous to mammalian Sp1 was cloned and inserted into a pET43.1a vector and expressed in Escherichia coli Rosetta (DE3) cells as a Nus-His-tag fusion protein. After induction with isopropyl thiogalactoside, the protein was purified with a Ni-Sepharose column, and approximately 5-8 mg of pure protein was obtained per liter of culture. The primary sequence and the predicted partial tertiary structure of the potential recombinant zebrafish Sp1 protein are similar to those of human Sp1. The DNA affinity precipitation assay and dual-luciferase promoter activity assay further confirm the nature of the recombinant zebrafish Sp1 protein as a transcription factor. Our results show that zebrafish Sp1-like protein is structurally and functionally comparable to human Sp1.",
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Zebrafish Sp1-like protein is structurally and functionally comparable to human Sp1. / Lin, Cha Jang; Hsiao, Tsun Hsien; Chung, Yi Shao; Chang, Wen Ni; Yeh, Trai-Ming; Chen, Bing Hung; Fu, Tzu-Fun.

In: Protein Expression and Purification, Vol. 76, No. 1, 01.03.2011, p. 36-43.

Research output: Contribution to journalArticle

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