Molecular cloning and characterization of canine Pre-B-Cell colony-enhancing factor

James R. McGlothlin, Li Gao, Tera Lavoie, Brett A. Simon, R. Blaine Easley, Shwu Fan Ma, Bernice B. Rumala, Joe GN Garcia, Shui Qing Ye

Research output: Contribution to journalArticle

70 Citations (Scopus)

Abstract

During our previous attempt to search for the candidate genes to acute lung injury (ALI), we unexpectedly identified PBEF as the most highly upregulated gene in a canine model of ALI by crosshybridizing canine lung cRNA to the Affymetrix human gene chip HG-U133A. The result suggested that PBEF may be a potential biomarker in ALI. To extend and translate that finding, we have performed the molecular cloning and characterization of canine PBEF cDNA in this study. Deduced amino acid sequence alignment revealed that the PBEF gene is evolutionarily highly conserved, with the canine PBEF protein sequence 96% identical to human PBEF and 94% identical to both murine and rat PBEF counterparts. Canine PBEF protein was successfully expressed both by in vitro transcription coupled with translation in a cell-free system and by transfection of canine PBEF cDNA into the human lung type II alveolar adenocarcinoma cell line A549. The expressed canine PBEF protein was visualized by either an anti-V5 tag peptide polyclonal antibody or an anti-canine PBEF peptide polyclonal antibody. RT-PCR assay indicates that canine PBEF is expressed in canine lung, brain, heart, liver, spleen, kidney, pancreas, and muscle, with liver showing the highest expression, followed by muscle. Isolation of the canine PBEF cDNA and expression of its recombinant protein may provide molecular tools to study the molecular mechanism of ALI in the canine model and to elucidate the potential role of PBEF as an ALI biomarker.

Original languageEnglish (US)
Pages (from-to)127-141
Number of pages15
JournalBiochemical Genetics
Volume43
Issue number3-4
DOIs
StatePublished - Apr 2005
Externally publishedYes

Fingerprint

Nicotinamide Phosphoribosyltransferase
Cloning
Molecular Cloning
B-lymphocytes
Canidae
molecular cloning
Genes
Complementary DNA
dogs
Biomarkers
lungs
Acute Lung Injury
protein
Liver
gene
Muscle
peptide
antibody
biomarker
muscle

Keywords

  • Canine
  • cDNA
  • Cloning
  • Gene expression
  • PBEF

ASJC Scopus subject areas

  • Biochemistry
  • Genetics

Cite this

McGlothlin, J. R., Gao, L., Lavoie, T., Simon, B. A., Easley, R. B., Ma, S. F., ... Ye, S. Q. (2005). Molecular cloning and characterization of canine Pre-B-Cell colony-enhancing factor. Biochemical Genetics, 43(3-4), 127-141. https://doi.org/10.1007/s10528-005-1505-2

Molecular cloning and characterization of canine Pre-B-Cell colony-enhancing factor. / McGlothlin, James R.; Gao, Li; Lavoie, Tera; Simon, Brett A.; Easley, R. Blaine; Ma, Shwu Fan; Rumala, Bernice B.; Garcia, Joe GN; Ye, Shui Qing.

In: Biochemical Genetics, Vol. 43, No. 3-4, 04.2005, p. 127-141.

Research output: Contribution to journalArticle

McGlothlin, JR, Gao, L, Lavoie, T, Simon, BA, Easley, RB, Ma, SF, Rumala, BB, Garcia, JGN & Ye, SQ 2005, 'Molecular cloning and characterization of canine Pre-B-Cell colony-enhancing factor', Biochemical Genetics, vol. 43, no. 3-4, pp. 127-141. https://doi.org/10.1007/s10528-005-1505-2
McGlothlin JR, Gao L, Lavoie T, Simon BA, Easley RB, Ma SF et al. Molecular cloning and characterization of canine Pre-B-Cell colony-enhancing factor. Biochemical Genetics. 2005 Apr;43(3-4):127-141. https://doi.org/10.1007/s10528-005-1505-2
McGlothlin, James R. ; Gao, Li ; Lavoie, Tera ; Simon, Brett A. ; Easley, R. Blaine ; Ma, Shwu Fan ; Rumala, Bernice B. ; Garcia, Joe GN ; Ye, Shui Qing. / Molecular cloning and characterization of canine Pre-B-Cell colony-enhancing factor. In: Biochemical Genetics. 2005 ; Vol. 43, No. 3-4. pp. 127-141.
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