Cloning and expression of murine interleukin-1 cDNA in Escherichia coli

Abstract

Interleukin-1 (IL-1), a peptide hormone produced by activated macrophages, possesses the ability to modulate the proliferation, maturation and functional activation of a broad spectrum of cell types and may play a major role in the initiation and amplification of immune and inflammatory responses through its action on these diverse cell populations. IL-1 exhibits microheterogeneity in terms of its relative molecular mass (Mr, 13,000-19,000) and charge properties, and although murine IL-1 has been purified and some of its basic structure-function relationships have been elucidated, it has proved difficult to prepare sufficient amounts of IL-1 for direct and detailed sequence and structural studies. Here we report the cloning, sequence analysis and expression of murine IL-1 cDNA in Escherichia coli. The IL-1 cDNA codes for a polypeptide precursor of 270 amino acids. Biologically active IL-1 was produced in E. coli by expressing the carboxy-terminal 156 amino acids of the IL-1 precursor.

Expression of interleukin-1 alpha, tumor necrosis factor alpha and interleukin-6 genes in astrocytes under ischemic injury

“…Fig. 5(A) showed the alignment between the cloned PCR product ampli®ed by the IL-1a primers and the mouse IL-1a cDNA (Lomedico et al, 1984; Genebank accession number: X01450). The extremely high homology indicated that the fragment was ampli®ed from IL-1a cDNA.…”

Regulation of the plasminogen activator inhibitor-2 (PAI-2) gene in murine macrophages. Demonstration of a novel pattern of responsiveness to bacterial endotoxin

“…26]; uPA; 1.2-kb PstI fragment of murine cDNA from pDB15 [29]; murine IL-1␤ [30]; ␤-actin, a 1.8-kb chicken ␤-actin cDNA fragment in pGEM2; c-fms, p755 containing the 3.7-kb mouse c-fms cDNA, obtained from L. Rohrschneider (Fred Hutchinson Cancer Research Centre, Seattle, WA); and lysozyme, pMLC, which contains a 780-bp fragment of mouse lysozyme M cDNA, obtained from R. Renkawitz (Max Planck Institute, Munich, Germany). Blots were reprobed with a synthetic 24-bp oligonucleotide (5Ј-CAT GGT AGG CAC GGC GAC TAC CAT-3Ј) complementary to murine 18S rRNA to control for gel loading.…”

A CRE/ATF-like site in the upstream regulatory sequence of the human interleukin 1 beta gene is necessary for induction in U937 and THP-1 monocytic cell lines.

“…The IL-1 cytokines consist of two polypeptides, IL-la and IL-1,B, that have essentially the same activity although they are the products of two different genes (4,30,31). The two polypeptides are produced as 31-kDa precursors which are subsequently processed to their 17-kDa active form (3,17).…”

“…These cytokines can act individually or in concert to exert an influence on a target cell or tissue (reviewed in reference 2). The IL-1 cytokines consist of two polypeptides, IL-la and IL-1,B, that have essentially the same activity although they are the products of two different genes (4,30,31 …”