Cloning and expression of caprine interferon-gamma

被引：32

作者：

Beyer, JC ^{[1
]}

Stich, RW ^{[1
]}

Hoover, DS ^{[1
]}

Brown, WC ^{[1
]}

Cheevers, WP ^{[1
]}

机构：

[1] Washington State Univ, Dept Vet Microbiol & Pathol, Pullman, WA 99164 USA

来源：

GENE | 1998年 / 210卷 / 01期

关键词：

caprine arthritis-encephalitis virus; plasmid vaccine; cytokine; expression plasmid;

D O I：

10.1016/S0378-1119(98)00046-8

中图分类号：

Q3 [遗传学];

学科分类号：

071007 ; 090102 ;

摘要：

Caprine interferon-gamma (IFN-gamma) cDNA was cloned from mitogen stimulated peripheral blood mononuclear cell (PBMC) RNA utilizing the reverse transcription-polymerase chain reaction (RT-PCR). The cDNA open reading frame (ORF) is 498 bp, encoding a putative 166 amino acid (aa) protein (19 327 Da). The predicted aa sequence homology of caprine IFN-gamma and the corresponding ovine, bovine and cervine cytokine is 98.8%, 95.2% and 92.8%, respectively. IFN-gamma cDNA was subcloned and expressed in two different plasmids under the control of either the human cytomegalovirus (CMV) immediate early promoter or the caprine arthritis-encephalitis virus long terminal repeat (CAEV LTR). Recombinant caprine IFN-gamma (rCaIFN-gamma) secreted by transfected COS-7 cells shared at least two antigenic epitopes with recombinant bovine IFN-gamma (rBoIFN-gamma) and exhibited biological activity in the vesicular stomatitis virus (VSV) cytopathic effect reduction assay. In-vivo expression of IFN-gamma cDNA promoted by the CAEV LTR was confirmed by the intramuscular (IM) injection of Balb/C mice with plasmid followed by Western blot analysis of mouse serum against purified rCaIFN-gamma produced in E. coli. (C) 1998 Elsevier Science B.V.

引用

页码：103 / 108

页数：6

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