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

Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology
Yau-Hung ChenHuai-Jen Tsai

Abstract

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 Ni(2+)-NTA column, we purified approximately 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.

References

Jan 25, 1992·Nucleic Acids Research·L HuynenA R Bellamy
Mar 11, 1991·Nucleic Acids Research·S H Pearson-White
Dec 1, 1989·Genes & Development·S J Rhodes, S F Konieczny
Jul 1, 1995·Developmental Biology·D F PinneyC P Emerson
Jun 21, 1994·Biochimica Et Biophysica Acta·P Y RescanB Fauconneau
Apr 12, 1996·The Journal of Biological Chemistry·P J CzernikB K Hurlburt
Oct 30, 1996·Molecular and Biochemical Parasitology·B ConnollyD F Smith
Apr 1, 1996·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·P Y Rescan, L Gauvry
Feb 1, 1997·Protein Expression and Purification·S J Maleki, B K Hurlburt
Sep 25, 1999·Molecular Reproduction and Development·C J HuangH J Tsai
Sep 26, 2000·Developmental Dynamics : an Official Publication of the American Association of Anatomists·Y XuZ Gong
Nov 18, 2000·Nature·C NeytP D Currie
Jan 11, 2000·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Y H ChenH J Tsai
Jan 3, 2001·Journal of Biochemistry·M KajiharaK Yasuda
Jan 3, 2001·Genesis : the Journal of Genetics and Development·Y H ChenH J Tsai
Jul 27, 2001·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·P Y Rescan

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Citations

Oct 9, 2002·Differentiation; Research in Biological Diversity·Yau-Hung Chen, Huai-Jen Tsai
May 21, 2009·Comparative Biochemistry and Physiology. Part B, Biochemistry & Molecular Biology·Øivind AndersenTrina Falck Galloway

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