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首页> 外文期刊>Cytokine >Porcine interleukin 18: cloning, characterization of the cDNA and expression with the baculovirus system.
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Porcine interleukin 18: cloning, characterization of the cDNA and expression with the baculovirus system.

机译：猪白介素18：杆状病毒系统的克隆，cDNA表征和表达。

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We have isolated and sequenced a cDNA that contains the coding sequence of porcine interleukin 18 (IL-18) and the recombinant protein of porcine IL-18 was expressed using the baculovirus system. The open reading frame (ORF) of the porcine IL-18 cDNA is 579 base pairs (bp) in length and encodes 192 amino acids. The predicted amino acid sequence is 76.7%, 64.7% and 61.6% homologous to the predicted human, murine and rat amino acid sequences, respectively. The porcine precursor and mature IL-18 protein were expressed respectively in Trichoplusia ni -derived (Tn5) cells using the baculovirus Autografha californica nuclear polyhedorosis virus (AcNPV) as a vector. Tn5 cells infected with recombinant virus containing a whole IL-18 protein coding region sequence secreted porcine precursor IL-18 into the culture medium. On the other hand, Tn5 cells infected with recombinant virus containing a mature IL-18 protein coding region sequence expressed several proteins in the cell lysates, but did not secrete mature protein into the culture medium efficiently. Immunoblotting analysis of recombinant protein showed cross-reactivity with anti-human IL-18 polyclonal antibody. The mature form of porcine IL-18 protein induced IFN-gamma production in suboptimal doses of anti-CD3 antibody and concanavalin A- (ConA) stimulated porcine peripheral blood mononuclear cells (PBMC), but the precursor form had little effect. Copyright 2000 Academic Press.
机译：我们已经分离并测序了包含猪白介素18（IL-18）编码序列的cDNA，并使用杆状病毒系统表达了猪IL-18的重组蛋白。猪IL-18 cDNA的开放阅读框（ORF）长度为579个碱基对（bp），编码192个氨基酸。预测的氨基酸序列分别与预测的人，鼠和大鼠氨基酸序列同源性为76.7％，64.7％和61.6％。使用杆状病毒Autografha californica核多角体病病毒（AcNPV）作为载体，分别在源自Trichoplusia ni的（Tn5）细胞中表达猪前体和成熟IL-18蛋白。用含有完整IL-18蛋白编码区序列的重组病毒感染的Tn5细胞将猪前体IL-18分泌到培养基中。另一方面，用含有成熟IL-18蛋白编码区序列的重组病毒感染的Tn5细胞在细胞裂解物中表达了几种蛋白，但没有有效地将成熟蛋白分泌到培养基中。重组蛋白的免疫印迹分析显示与抗人IL-18多克隆抗体具有交叉反应性。猪IL-18蛋白的成熟形式在次最佳剂量的抗CD3抗体和伴刀豆球蛋白A-（ConA）刺激的猪外周血单个核细胞（PBMC）中诱导IFN-γ产生，但前体形式几乎没有作用。版权所有2000学术出版社。

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