Devlin J J, Devlin P E, Myambo K, Lilly M B, Rado T A, Warren M K
J Leukoc Biol. 1987 Apr;41(4):302-6. doi: 10.1002/jlb.41.4.302.
We have isolated and expressed a cDNA clone that encodes a human granulocyte colony-stimulating factor from the MIA PaCa-2 cell line. A genomic clone of this factor has been isolated from the CHU-2 cell line and is reported to encode two alternative transcripts [The EMBO J. 5,575, 1986]; one transcript predicts an amino acid sequence identical to that predicted by our MIA PaCa-2 cDNA clone; the other transcript predicts a similar protein containing a three amino acid residue insertion. To investigate which types of this colony-stimulating factor are produced by other cell lines, we used specific oligonucleotides to determine which types of transcripts were present in MIA PaCa-2, 5637, and LD-1 cells, all of which have been reported to produce a factor that can stimulate the growth of predominantly granulocyte colonies in human bone marrow cell cultures. Northern analysis with these probes revealed MIA PaCa-2-like transcripts in all of these cell lines and failed to detect transcripts that would encode the colony-stimulating factor that contained the three-amino-acid-residue insertion.
我们从MIA PaCa - 2细胞系中分离并表达了一个编码人粒细胞集落刺激因子的cDNA克隆。该因子的一个基因组克隆已从CHU - 2细胞系中分离出来,据报道它编码两种可变转录本[《欧洲分子生物学组织杂志》5,575, 1986年];一种转录本预测的氨基酸序列与我们的MIA PaCa - 2 cDNA克隆预测的序列相同;另一种转录本预测的是一种相似的蛋白质,含有一个三氨基酸残基插入。为了研究其他细胞系产生哪种类型的这种集落刺激因子,我们使用特异性寡核苷酸来确定MIA PaCa - 2、5637和LD - 1细胞系中存在哪种类型的转录本,所有这些细胞系都已被报道能产生一种可刺激人骨髓细胞培养物中主要是粒细胞集落生长的因子。用这些探针进行的Northern分析在所有这些细胞系中都检测到了MIA PaCa - 2样转录本,并且未检测到编码含有三氨基酸残基插入的集落刺激因子的转录本。