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 journalArticlepeer-review

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

All Science Journal Classification (ASJC) codes

  • Biotechnology

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