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Title: Cloning and expression of chicken granulocyte-macrophage colony stimulating factor (GMCSF) gene
Authors: Chaturvedi, Uttara
Kalim, Shahina
Kumar, Rajiv
Sawant, Pradeep
Tiwari, Sangeeta
Khurana, S K
Sahoo, A P
Palia, Sudesh
Tiwari, Ashok Kumar
Keywords: Genetic adjuvant;GMCSF;PCR
Issue Date: Dec-2010
Publisher: NISCAIR-CSIR, India
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.
Page(s): 1175-1180
ISSN: 0975-1009 (Online); 0019-5189 (Print)
Appears in Collections:IJEB Vol.48(12) [December 2010]

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