Back to Search
Start Over
Cloning of porcine interleukin (IL)-12 receptor beta2 (IL-12Rbeta2) gene and its application to a rapid biological assay for human/porcine IL-12
- Source :
- Veterinary immunology and immunopathology. 91(2)
- Publication Year :
- 2003
-
Abstract
- Porcine IL-12Rbeta2 gene was cloned from mRNA preparation of mitogen-activated peripheral blood mononuclear cells (PBMCs), and its complete nucleotide sequence was determined. To confirm the biological function, the entire open reading frame (ORF) was re-cloned into a mammalian expression vector, pcDNA3.1/Zeo(+), at the downstream of CMV promoter, and introduced to a Th1-like human lymphoma cell line, Jurkat E6-1. Antibiotic-resistant cells retaining the expression construct were selected then, isolated by the limiting dilution method. An established clone (10B10) constitutively expressed chimeric IL-12Rs composed of intrinsic (human) beta1 and extrinsic (porcine) beta2 subunits, and produced interferon (IFN)-gamma in response to IL-12 of both species with optimal PHA/PMA stimulation. The production of IFN-gamma was observed as early as 42 h after culture and appeared to be dose-dependent within the range between 20 and 2000 pg/ml. Thus, this clone not only reacts with IL-12 of both species but also provides a useful tool for quick and sensitive detection of IL-12 bioactivity.
- Subjects :
- Swine
Immunology
Molecular Sequence Data
Clone (cell biology)
Gene Expression
Biology
Jurkat cells
Peripheral blood mononuclear cell
Interferon
medicine
Animals
Humans
Amino Acid Sequence
RNA, Messenger
Cloning, Molecular
Expression vector
General Veterinary
Sequence Homology, Amino Acid
Receptors, Interleukin-12
Receptors, Interleukin
Molecular biology
Interleukin-12
Open reading frame
Cell culture
Interleukin 12
medicine.drug
Subjects
Details
- ISSN :
- 01652427
- Volume :
- 91
- Issue :
- 2
- Database :
- OpenAIRE
- Journal :
- Veterinary immunology and immunopathology
- Accession number :
- edsair.doi.dedup.....aaa827be26d472f2b82d3c0ce77e9e23