Molecular cloning and expression of the cDNA encoding feline granulocyte colony-stimulating factor.
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.
Yamamoto
A
A
Nippon Institute for Biological Science, Shin-Machi 9-2221-1, Ome, Tokyo 198-0024, Japan. yamagen@mx6.ttcn.ne.jp
Iwata
A
A
Tuchiya
K
K
Katsumata
A
A
Oishi
K
K
Saito
T
T
Tsujimoto
H
H
Hasegawa
A
A
Ueda
S
S
D000595:Amino Acid Sequence; D000818:Animals; D002415:Cats; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000235:genetics; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics*; D004305:Dose-Response Relationship, Drug; D005091:Exons; D015870:Gene Expression; D005796:Genes / Q000235:genetics; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics* / Q000494:pharmacology; D006801:Humans; D007438:Introns; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D012333:RNA, Messenger; D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D004247:DNA