Cloning and expression of the cervine interleukin 4 gene.
Interleukin 4 (IL-4), a cytokine produced by the T helper 2 subset of T lymphocytes is involved in up regulating antibody responses. A source of recombinant cervine IL-4 would be useful for studying the immune response of deer to tuberculosis. We report here the cloning, sequencing and expression of recombinant cervine IL-4. To achieve this mRNA was isolated from red deer (Cervus elaphus) mononuclear leucocytes. First strand cDNA was synthesized from the mRNA and the IL-4 cDNA was amplified, cloned and sequenced. The IL-4 cDNA is 408 bp in length and the deduced amino acid sequence is 92% homologous with the published bovine IL-4 amino acid sequence. IL-4 was expressed using the baculovirus expression system in Spodoptera frugiperda Sf9 cells. Northern blot, SDS-PAGE analysis and bioassay were used to confirm the expression of IL-4.
Deer Research Laboratory, Department of Microbiology, University of Otago, Dunedin, New Zealand.
S M
SM
Hook
A M
AM
Crawford
D N
DN
Chinn
J F
JF
Griffin
G S
GS
Buchan
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001681:Biological Assay; D015152:Blotting, Northern; D003001:Cloning, Molecular; D004274:DNA, Recombinant; D003670:Deer / Q000235:genetics / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D015847:Interleukin-4 / Q000096:biosynthesis* / Q000235:genetics*; D008213:Lymphocyte Activation; D008969:Molecular Sequence Data; D009036:Moths; D017924:Nucleopolyhedroviruses / Q000235:genetics; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000096:biosynthesis; D017386:Sequence Homology, Amino Acid
D004274:DNA, Recombinant; D011994:Recombinant Proteins; D015847:Interleukin-4