Cloning and expression of chicken granulocyte-macrophage colony stimulating factor (GMCSF) gene.
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.
Molecular Biology Laboratory, Department of Animal Biotechnology, Indian Veterinary Research Institute, Izatnagar 243 122, UP, India.
Uttara
U
Chaturvedi
Shahina
S
Kalim
Rajiv
R
Kumar
Pradeep
P
Sawant
Sangeeta
S
Tiwari
S K
SK
Khurana
A P
AP
Sahoo
Sudesh
S
Palia
Ashok Kumar
AK
Tiwari
D000818:Animals; D001483:Base Sequence; D002645:Chickens; D002522:Chlorocebus aethiops; D003001:Cloning, Molecular; D005822:Genetic Vectors; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics* / Q000378:metabolism*; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012689:Sequence Homology, Nucleic Acid; D014709:Vero Cells
D012333:RNA, Messenger; D016178:Granulocyte-Macrophage Colony-Stimulating Factor