Cloning and expression of chicken granulocyte-macrophage colony stimulating factor (GMCSF) gene.
Loading...
Date
2010-12
Journal Title
Journal ISSN
Volume Title
Publisher
Abstract
Granulocyte-macrophage colony stimulating factor (GMCSF), a multifunctional cytokine can enhance immune responses when administered along with DNA vaccine. Aim of the present study was to clone and express the chicken GMCSF cytokine for use as ‘genetic adjuvant’. Chicken GMCSF gene 435bp was amplified using specific primers in which restriction sites of BamHI and HindIII were at forward and reverse primers respectively. The PCR product was cloned into eukaryotic expression vector pcDNA 3.1(+) and clones were confirmed by restriction digestion and nucleotide sequencing. Functional activity of recombinant GMCSF was checked by expression of GMCSF specific mRNA in transfected Vero cells by RT-PCR of total RNA isolated from transfected Vero cells. The recombinant plasmid can be used as genetic adjuvant in chicken.
Description
Keywords
Genetic adjuvant, GMCSF, PCR
Citation
Chaturvedi Uttara, Kalim Shahina, Kumar Rajiv, Sawant Pradeep, Tiwari Sangeeta, Khurana S K, Sahoo A P, Palia Sudesh, Tiwari Ashok Kumar. Cloning and expression of chicken granulocyte-macrophage colony stimulating factor (GMCSF) gene. Indian Journal of Experimental Biology. 2010 Dec; 48(12): 1175-1180.