Bayne M L, Cascieri M A, Kelder B, Applebaum J, Chicchi G, Shapiro J A, Pasleau F, Kopchick J J
Proc Natl Acad Sci U S A. 1987 May;84(9):2638-42. doi: 10.1073/pnas.84.9.2638.
A synthetic gene encoding human insulin-like growth factor I (hIGF-I) was assembled and inserted into an expression vector containing the cytomegalovirus immediate early (CMV-IE) transcriptional regulatory region and portions of the bovine growth hormone gene. The recombinant plasmid encodes a 97 amino acid fusion protein containing the first 27 amino acids of the bovine growth hormone precursor and the 70 amino acids of hIGF-I. This plasmid, when transiently introduced into cultured mouse fibroblasts, directs synthesis of the fusion protein, subsequent proteolytic removal of the bovine growth hormone signal peptide, and secretion of hIGF-I into the culture medium. Conditioned medium from transfected cells inhibits binding of 125I-labeled IGF-I to type I IGF receptors on human placental membranes and to acid-stable human serum carrier proteins. The recombinant hIGF-I produced is biologically active, as monitored by the stimulation of DNA synthesis in vascular smooth muscle cells.
一个编码人胰岛素样生长因子I(hIGF-I)的合成基因被组装并插入到一个表达载体中,该表达载体包含巨细胞病毒立即早期(CMV-IE)转录调控区和牛生长激素基因的部分序列。重组质粒编码一种97个氨基酸的融合蛋白,其中包含牛生长激素前体的前27个氨基酸和hIGF-I的70个氨基酸。该质粒在瞬时导入培养的小鼠成纤维细胞后,指导融合蛋白的合成,随后通过蛋白水解去除牛生长激素信号肽,并将hIGF-I分泌到培养基中。转染细胞的条件培养基可抑制125I标记的IGF-I与人胎盘膜上的I型IGF受体以及酸稳定的人血清载体蛋白的结合。通过刺激血管平滑肌细胞中的DNA合成监测,所产生的重组hIGF-I具有生物活性。