Cloning of equine interleukin 1 receptor antagonist and determination of its full-length cDNA sequence

Rick D. Howard From the Orthopaedic Research Laboratory, Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences (Howard, McIlwraith, Trotter) and the Department of Biochemistry and Molecular Biology, College of Natural Sciences (Nyborg), Colorado State University, Fort Collins, CO 80523.

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 DVM, PhD
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C.Wayne McIlwraith From the Orthopaedic Research Laboratory, Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences (Howard, McIlwraith, Trotter) and the Department of Biochemistry and Molecular Biology, College of Natural Sciences (Nyborg), Colorado State University, Fort Collins, CO 80523.

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Gayle W. Trotter From the Orthopaedic Research Laboratory, Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences (Howard, McIlwraith, Trotter) and the Department of Biochemistry and Molecular Biology, College of Natural Sciences (Nyborg), Colorado State University, Fort Collins, CO 80523.

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 DVM, MS
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Jennifer K. Nyborg From the Orthopaedic Research Laboratory, Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences (Howard, McIlwraith, Trotter) and the Department of Biochemistry and Molecular Biology, College of Natural Sciences (Nyborg), Colorado State University, Fort Collins, CO 80523.

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 PhD

Abstract

Objectives

To clone equine interleukin 1 receptor antagonist (IL-1 ra) and determine its full-length cDNA sequence.

Procedure

A cDNA library derived from lipopolysaccharide-stimulated equine monocytes was screened by means of plaque hybridization to radiolabeled equine IL-1ra DNA probes generated by means of the polymerase chain reaction. The cDNA nucleotide sequence for equine IL-1ra was determined by use of the dideoxy chain termination technique, analyzed by use of computer software for sequence characteristics, and compared with sequences reported for IL-1ra of other species.

Results

The cDNA for equine IL-1ra was 1,614 base pairs in length with an ORF encoding a peptide of 177 amino acids with a predicted molecular mass of 20.427 kd. Similarity between the amino acid sequence of equine IL-1ra and sequences for human, murine, rat, and lapine IL-1ra was 76%. Similarity between sequence for equine IL-1ra and sequences for equine interleukin-1α and equine interleukin-1ß were 22.6 and 24.6%, respectively.

Conclusion

Comparison of the sequence for equine IL-1ra with sequences for IL-1ra of other species indicated a high degree of conservation.

Clinical Relevance

Results establish a basis for studying the roles of interleukin-1 in healthy and diseased joints in horses. (Am J Vet Res 1998;59:712-716)

Abstract

Objectives

To clone equine interleukin 1 receptor antagonist (IL-1 ra) and determine its full-length cDNA sequence.

Procedure

A cDNA library derived from lipopolysaccharide-stimulated equine monocytes was screened by means of plaque hybridization to radiolabeled equine IL-1ra DNA probes generated by means of the polymerase chain reaction. The cDNA nucleotide sequence for equine IL-1ra was determined by use of the dideoxy chain termination technique, analyzed by use of computer software for sequence characteristics, and compared with sequences reported for IL-1ra of other species.

Results

The cDNA for equine IL-1ra was 1,614 base pairs in length with an ORF encoding a peptide of 177 amino acids with a predicted molecular mass of 20.427 kd. Similarity between the amino acid sequence of equine IL-1ra and sequences for human, murine, rat, and lapine IL-1ra was 76%. Similarity between sequence for equine IL-1ra and sequences for equine interleukin-1α and equine interleukin-1ß were 22.6 and 24.6%, respectively.

Conclusion

Comparison of the sequence for equine IL-1ra with sequences for IL-1ra of other species indicated a high degree of conservation.

Clinical Relevance

Results establish a basis for studying the roles of interleukin-1 in healthy and diseased joints in horses. (Am J Vet Res 1998;59:712-716)

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