鹿白细胞介素4基因的克隆与表达

Cloning and expression of the cervine interleukin 4 gene.

作者信息

Hook S M, Crawford A M, Chinn D N, Griffin J F, Buchan G S

机构信息

Deer Research Laboratory, Department of Microbiology, University of Otago, Dunedin, New Zealand.

出版信息

Scand J Immunol. 1994 Jul;40(1):71-6. doi: 10.1111/j.1365-3083.1994.tb03435.x.

DOI:10.1111/j.1365-3083.1994.tb03435.x

PMID:8029646

Abstract

Interleukin 4 (IL-4), a cytokine produced by the T helper 2 subset of T lymphocytes is involved in up regulating antibody responses. A source of recombinant cervine IL-4 would be useful for studying the immune response of deer to tuberculosis. We report here the cloning, sequencing and expression of recombinant cervine IL-4. To achieve this mRNA was isolated from red deer (Cervus elaphus) mononuclear leucocytes. First strand cDNA was synthesized from the mRNA and the IL-4 cDNA was amplified, cloned and sequenced. The IL-4 cDNA is 408 bp in length and the deduced amino acid sequence is 92% homologous with the published bovine IL-4 amino acid sequence. IL-4 was expressed using the baculovirus expression system in Spodoptera frugiperda Sf9 cells. Northern blot, SDS-PAGE analysis and bioassay were used to confirm the expression of IL-4.

摘要

白细胞介素4（IL-4）是由T淋巴细胞的辅助性T细胞2亚群产生的一种细胞因子，参与上调抗体反应。重组鹿IL-4来源对于研究鹿对结核病的免疫反应将很有用。我们在此报告重组鹿IL-4的克隆、测序和表达。为实现此目的，从马鹿（Cervus elaphus）单核白细胞中分离出mRNA。从mRNA合成第一链cDNA，并对IL-4 cDNA进行扩增、克隆和测序。IL-4 cDNA长度为408 bp，推导的氨基酸序列与已发表的牛IL-4氨基酸序列有92%的同源性。利用杆状病毒表达系统在草地贪夜蛾Sf9细胞中表达IL-4。采用Northern印迹、SDS-PAGE分析和生物测定法来确认IL-4的表达。