Research Article

Cloning, expression analysis and sequence prediction of the CCAAT/enhancer-binding protein alpha gene of Qinchuan cattle

Published: June 15, 2012
Genet. Mol. Res. 11 (2) : 1651-1661 DOI: https://doi.org/10.4238/2012.June.15.14
Cite this Article:
(2012). Cloning, expression analysis and sequence prediction of the CCAAT/enhancer-binding protein alpha gene of Qinchuan cattle. Genet. Mol. Res. 11(2): gmr1567. https://doi.org/10.4238/2012.June.15.14
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Abstract

CCAAT/enhancer-binding protein alpha (C/EBPα) is an essential transcription factor, regulating the differentiation of adipocytes. We cloned the complete open reading frame of C/EBPα gene of Qinchuan cattle and analyzed its protein structures and expression profile in 15 tissues via DNA cloning, sequencing and RT-PCR. Analysis of the putative protein sequences revealed that C/EBPα consists of alpha helices, random coils and a few extended strands. A significant transmembrane structure was observed in amino acid region 233 to 252. A basic leucine zipper domain was also found in amino acid region 277 to 340, which is characteristic of C/EBPs. Homologous comparison with various species indicated that the C/EBPα gene of Qinchuan cattle shares 97, 95, 94, 94, and 93% similarity in amino acid sequences with Sus scrofa, Homo sapiens, Rattus norvegicus, Oryctolagus cuniculus, and Mus musculus, respectively, implying strong sequence conservation of C/EBPα during evolution. RT-PCR revealed that the mRNA expression level of bovine C/EBPα gene in subcutaneous fat is much higher than that in the other 14 tissues, and the relative quantity in fat tissue increases with cattle age.

CCAAT/enhancer-binding protein alpha (C/EBPα) is an essential transcription factor, regulating the differentiation of adipocytes. We cloned the complete open reading frame of C/EBPα gene of Qinchuan cattle and analyzed its protein structures and expression profile in 15 tissues via DNA cloning, sequencing and RT-PCR. Analysis of the putative protein sequences revealed that C/EBPα consists of alpha helices, random coils and a few extended strands. A significant transmembrane structure was observed in amino acid region 233 to 252. A basic leucine zipper domain was also found in amino acid region 277 to 340, which is characteristic of C/EBPs. Homologous comparison with various species indicated that the C/EBPα gene of Qinchuan cattle shares 97, 95, 94, 94, and 93% similarity in amino acid sequences with Sus scrofa, Homo sapiens, Rattus norvegicus, Oryctolagus cuniculus, and Mus musculus, respectively, implying strong sequence conservation of C/EBPα during evolution. RT-PCR revealed that the mRNA expression level of bovine C/EBPα gene in subcutaneous fat is much higher than that in the other 14 tissues, and the relative quantity in fat tissue increases with cattle age.