Cloning and production of hamster recombinant INFG antigen for production of anti-INFG antibodies (2006)
- Authors:
- Autor USP: GIDLUND, MAGNUS AKE - ICB
- Unidade: ICB
- Assunto: IMUNOLOGIA
- Language: Inglês
- Abstract: Introduction: Interferon gamma (IFN g) is an imunomodulatory citokyne produced in inflammation. It is associated with several pathologic processes including infectious diseases. Several studies have associated IFN g with atherosclerotic process in mice, rabbits and humans. Golden hamster (Mesocricetus auratus) was widely used as a model for atherosclerosis because of its lipid metabolism that is similar to humans, however the model is not widely used because we lack immunological reagents. Therefore, the need for such reagents has to be solved producing our own reagents to proceed the work with this animal model. Objective: The aim of this work was to clone part of INF g from Golden Syriam Hamster and produce recombinant antigen for further production of anti-INF g antibodies, aiming detection tests and characterization of inflammatory response in the hamster model of experimental atherosclerosis.Methods: Primers were designed with two restriction sites Bam HI and Hind III based on the sequence of INF g accessed from NCBI's Entrez Genome database ( "AAC53580,1").Total RNA was extracted with Trizol® Invitrogen from splenic cells of hamster infected with Leishmania (L) chagasi. The mRNA was reverse Transcripted,PCRamplified, cloned in pGEM-T Easy vector(Promega), sub-cloned in Pet 28 a(+) (Novagen), overexpressed in Escherichia coli and purified by affinity chromatography. Results : The recombinant antigen was expressed at the level of3 mg / liter of culture. Purified product was obtained with little contamination with other proteins. Only problem was the precipitation of antigen that precipitates in high concentrations (up to 1 mg/ml) and in freezing conditions. Conclusions: In this work we have obtained a good amount of purified recombinant antigen in the conditions to utilize directly in the immunization of mice for the production of antibody, without need of any further step of purification.
- Imprenta:
- Source:
- Título: Abstracts
- Conference titles: Meeting of the Brazilian Society for Immunology
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ABNT
SANCHEZ, E M R et al. Cloning and production of hamster recombinant INFG antigen for production of anti-INFG antibodies. 2006, Anais.. São Paulo: Instituto de Ciências Biomédicas, Universidade de São Paulo, 2006. . Acesso em: 20 mar. 2026. -
APA
Sanchez, E. M. R., Goto, H., Columbano, C. O., Godefeder, M. B., & Gidlund, M. A. (2006). Cloning and production of hamster recombinant INFG antigen for production of anti-INFG antibodies. In Abstracts. São Paulo: Instituto de Ciências Biomédicas, Universidade de São Paulo. -
NLM
Sanchez EMR, Goto H, Columbano CO, Godefeder MB, Gidlund MA. Cloning and production of hamster recombinant INFG antigen for production of anti-INFG antibodies. Abstracts. 2006 ;[citado 2026 mar. 20 ] -
Vancouver
Sanchez EMR, Goto H, Columbano CO, Godefeder MB, Gidlund MA. Cloning and production of hamster recombinant INFG antigen for production of anti-INFG antibodies. Abstracts. 2006 ;[citado 2026 mar. 20 ] - Opposing effects between short-term combined transdermal estradiol/noretisterone against oxidized LDL (oxLDL) and heat shock proteins (hsp)
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