Yamamoto A, Iwata A, Tuchiya K, Katsumata A, Oishi K, Saito T, Tsujimoto H, Hasegawa A, Ueda S
Nippon Institute for Biological Science, Shin-Machi 9-2221-1, Ome, Tokyo 198-0024, Japan.
Gene. 2001 Aug 22;274(1-2):263-9. doi: 10.1016/s0378-1119(01)00575-3.
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.
猫粒细胞集落刺激因子（G-CSF）基因的基因组DNA和cDNA均从CRFK细胞中克隆得到。Southern印迹分析表明，单倍体基因组包含G-CSF基因的单拷贝。对几种猫细胞系的RT-PCR分析显示，在脂多糖刺激下G-CSF mRNA表达。基因组和cDNA克隆的序列分析表明，人和猫G-CSF基因之间的内含子-外显子连接结构是保守的。G-CSF编码区编码一个预测的195个氨基酸的蛋白质，包括一个21个氨基酸的信号序列。猫G-CSF氨基酸序列与犬（90.8%）、人（87.4%）、绵羊（83.9%）、牛（82.8%）、猪（80.5%）、小鼠（70.7%）和大鼠（66.8%）的G-CSF具有高度同源性。使用重组杆状病毒载体在昆虫细胞中表达的猫G-CSF，在使用NFS-60细胞的增殖试验和猫白细胞诱导试验中测定具有生物活性。
