Cloning of equine interleukin 1α and equine interleukin 1ß and determination of their full-length cDNA sequences

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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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 1α (IL-1α) and equine interleukin 1β (IL-1β) and determine their full-length cDNA sequences.

Procedure

The mRNA isolated from lipopolysaccharide-stimulated cultured equine monocytes was reverse transcribed, and a cDNA library was constructed in a λ phage. The cDNA library was screened by means of plaque hybridization with radiolabeled human IL-1α and IL-1ß cDNA probes. The cDNA nucleotide sequences for equine IL-1α and equine IL-1β were determined by use of the dideoxy chain termination technique. The cDNA sequences were analyzed, using computer software, for sequence characteristics and compared with sequences reported for other species.

Results

The cDNA for equine IL-1α was 1,728 base pairs in length with an ORF encoding a peptide of 270 amino acids with a predicted molecular mass of 30.823 kd. The cDNA for equine IL-1β was 1,473 base pairs in length with an ORF encoding a peptide of 268 amino acids with a predicted molecular mass of 30.342 kd. Similarity between amino acid sequence of equine IL- 1α and sequences for IL-1 α of other species ranged from 62.5 to 82.2%; similarity between amino acid sequence of equine IL-1ß and sequences for IL-1β of other species ranged from 62.5 to 66.4%. Similarity between amino acid sequences of equine IL-1α and equine IL-1β was 26%.

Conclusions and 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;59:704-711)

Abstract

Objectives

To clone equine interleukin 1α (IL-1α) and equine interleukin 1β (IL-1β) and determine their full-length cDNA sequences.

Procedure

The mRNA isolated from lipopolysaccharide-stimulated cultured equine monocytes was reverse transcribed, and a cDNA library was constructed in a λ phage. The cDNA library was screened by means of plaque hybridization with radiolabeled human IL-1α and IL-1ß cDNA probes. The cDNA nucleotide sequences for equine IL-1α and equine IL-1β were determined by use of the dideoxy chain termination technique. The cDNA sequences were analyzed, using computer software, for sequence characteristics and compared with sequences reported for other species.

Results

The cDNA for equine IL-1α was 1,728 base pairs in length with an ORF encoding a peptide of 270 amino acids with a predicted molecular mass of 30.823 kd. The cDNA for equine IL-1β was 1,473 base pairs in length with an ORF encoding a peptide of 268 amino acids with a predicted molecular mass of 30.342 kd. Similarity between amino acid sequence of equine IL- 1α and sequences for IL-1 α of other species ranged from 62.5 to 82.2%; similarity between amino acid sequence of equine IL-1ß and sequences for IL-1β of other species ranged from 62.5 to 66.4%. Similarity between amino acid sequences of equine IL-1α and equine IL-1β was 26%.

Conclusions and 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;59:704-711)

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