| Autor: |
Wong EY; Department of Biological Sciences, Monsanto Co., Chesterfield, MO 63198., Seetharam R, Kotts CE, Heeren RA, Klein BK, Braford SR, Mathis KJ, Bishop BF, Siegel NR, Smith CE, et. al. |
| Jazyk: |
angličtina |
| Zdroj: |
Gene [Gene] 1988 Sep 07; Vol. 68 (2), pp. 193-203. |
| DOI: |
10.1016/0378-1119(88)90021-2 |
| Abstrakt: |
The synthesis, processing and secretion of insulin-like growth factor-1 (IGF-1 or somatomedin-C) fused to LamB and OmpF secretion leader sequences in Escherichia coli have been investigated. Expression and secretion of IGF-1 was achieved. The major portion of this secreted IGF-1 accumulated in the periplasmic space as insoluble aggregates. A small amount of IGF-1 was found folded in its native conformation in the medium. The lamB and ompF signal sequences were fused to the 5' coding sequence of IGF-1. Fusion of the lamB signal sequence directly to IGF-1 (lamB-IGF-1) resulted in accumulation of 16-20 micrograms/A550/ml of correctly processed IGF-1 in the periplasmic space. The processing efficiency of LamB-IGF-1 and OmpF-IGF-1 was enhanced in an E. coli strain bearing a prlA4 mutation. Amino acid sequence analysis of IGF-1 secreted into the periplasm and exported into the medium confirmed the precise removal of the LamB or OmpF signal sequence. IGF-1 synthesized in E. coli was demonstrated to be active in a cell proliferation bioassay. |
| Databáze: |
MEDLINE |
| Externí odkaz: |
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