Tweardy D J, Cannizzaro L A, Palumbo A P, Shane S, Huebner K, Vantuinen P, Ledbetter D H, Finan J B, Nowell P C, Rovera G
Wistar Institute of Anatomy and Biology, Philadelphia, Pennsylvania 19104.
Oncogene Res. 1987 Aug;1(3):209-20.
The conditioned media (CM) of the glioblastoma multiforme cell line, U87 MG, contains abundant granulocyte colony-stimulating factor (G-CSF) activity (Tweardy et al., 1987). An oligonucleotide encoding the amino acids -11 to -4 of G-CSF detected a single abundant G-CSF mRNA of 1.6 kilobases (Kb) produced by U87 MG cells. Screening of a U87 MG cDNA library with the oligonucleotide identified cDNA clones of 1.3-1.4 Kb. Sequencing of one clone (pG-CSF6) confirmed that it encoded G-CSF and was derived from G-CSFb mRNA encoding a protein with a three amino acid deletion at positions 36-38. Only a single base substitution was observed at the third position of the codon for leu 152 indicating that G-CSF is highly conserved in cells of widely different origin. Somatic cell hybridization studies and chromosomal in situ hybridization localized the G-CSF gene to the long arm of chromosome 17 in band 17q21, proximal to the 17q breakpoint characteristic of acute promyelocytic leukemia.
多形性胶质母细胞瘤细胞系U87 MG的条件培养基(CM)含有丰富的粒细胞集落刺激因子(G-CSF)活性(Tweardy等人,1987年)。编码G-CSF氨基酸-11至-4的寡核苷酸检测到U87 MG细胞产生的一条1.6千碱基(Kb)的丰富G-CSF mRNA。用该寡核苷酸筛选U87 MG cDNA文库,鉴定出1.3 - 1.4 Kb的cDNA克隆。对一个克隆(pG-CSF6)进行测序,证实其编码G-CSF,且来源于G-CSFb mRNA,该mRNA编码的蛋白质在第36 - 38位有三个氨基酸缺失。在亮氨酸152密码子的第三位仅观察到一个单碱基替换,表明G-CSF在来源广泛不同的细胞中高度保守。体细胞杂交研究和染色体原位杂交将G-CSF基因定位到17号染色体长臂17q21带,靠近急性早幼粒细胞白血病特征性的17q断点。