Molecular cloning and expression of the cDNA encoding feline granulocyte colony-stimulating factor. |
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Authors: | A Yamamoto A Iwata K Tuchiya A Katsumata K Oishi T Saito H Tsujimoto A Hasegawa S Ueda |
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Institution: | Nippon Institute for Biological Science, Shin-Machi 9-2221-1, Ome, Tokyo 198-0024, Japan. yamagen@mx6.ttcn.ne.jp |
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Abstract: | Both genomic DNA and cDNA of the feline granulocyte colony-stimulating factor (G-CSF) gene were cloned from CRFK cells. Southern blot analysis showed that the haploid genome contains a single copy of the G-CSF gene. The RT-PCR analysis of several feline cell lines revealed expression of G-CSF mRNA in response to lipopolysaccharide stimulation. Sequence analysis of genomic and cDNA clones indicated that the intron-exon junction structure is conserved between the human and the feline G-CSF genes. The G-CSF coding region encodes a predicted protein of 195 amino acids including a signal sequence of 21 amino acids. The feline G-CSF amino acid sequence shares a high degree of identity with the canine (90.8%), human (87.4%), ovine (83.9%), bovine (82.8%), porcine (80.5%), murine (70.7%) and rat (66.8%) G-CSF. The feline G-CSF expressed in insect cells using recombinant baculovirus vector was biologically active as measured in a proliferation assay using NFS-60 cells and an induction assay of leukocytes in cats. |
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