Volume 10, Issue 3 (vol- 3 2004)                   Horizon Med Sci 2004, 10(3): 20-30 | Back to browse issues page

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Azadmanesh K, Roohvand F, Amini S, Andalibi Mahmoodabadi S, Kazanji M. Isolation of IFN- gamma gene from olive baboon and designed semi-quantitative competitive PCR method to quantify gene expression. Horizon Med Sci. 2004; 10 (3) :20-30
URL: http://hms.gmu.ac.ir/article-1-274-en.html
1- , azadmanesh@pasteur.zc.ir
Abstract:   (13228 Views)
Background and Aim: Primates have long been used as the most resembling animal model to human. Primates are natural hosts of HTLV/STLV retroviruses. From this family, HTLV-I is a health problem in Khorasan. Recently a new HTLV-I resembling virus has been detected in olive baboon, which suggests this animal as an ideal animal model for therapeutic studies of this disease, for such studies inventing methods and tools to quantify cytokines of this animal, especially IFN-, has high priority. Materials and Methods: Here in, we report methods for isolating cDNA of IFN- from peripheral blood, comparison of its sequence with other primates, construction of competitive plasmids and primers for quantifying this cytokine. In order to standardize the results of quantification, we needed to measure the expression level of a house keeping gene too, so we designed a competitor plasmid and set of primers to quantify the expression of Glycer Aldehyde 3 Phosphate DeHydrogenase (G3PDH) too. Results: Two new cDNA sequences of baboon IFN- and G3PDH were registered in NCBI GENBANK. The competitive PCRs of a sample of this animal measured 7 pg and 0.455 pgof IFN- and G3PDH transcripts respectively. Conclusion: Phylogenetic analysis of IFN- well classified this baboon among old world monkeys, while the structure of this cytokine seems to be same as human. According to the methodology presented here, it is possible to use the ratio of IFN-/G3PDH expression as a tool to compare levels of IFN- production in different samples.
Full-Text [DOC 430 kb]   (3544 Downloads)    
Type of Study: Original | Subject: Internal Medicine
Received: 2008/07/26 | Published: 2004/10/15

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