| Autor: |
Arthur Hawtrey, Mario Ariatti, Barbara Huckett |
| Rok vydání: |
1990 |
| Předmět: |
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| Zdroj: |
Biochemical Pharmacology. 40:253-263 |
| ISSN: |
0006-2952 |
| DOI: |
10.1016/0006-2952(90)90686-f |
| Popis: |
Evidence is presented for targeted gene delivery to HepG2 cells via the endocytotic pathway under the direction of insulin. Serum albumin was treated with the water-soluble carbodiimide N -ethyl- N ′-(3-dimethylaminopropyl) carbodiimide hydrochloride and the resultant positively charged N -acylurea albumin covalently conjugated to insulin by glutaraldehyde cross-linkage. The conjugated protein is shown by nitrocellulose filter binding and gel band shift assays to bind DNA, and by competitive displacement of [ 125 I]insulin to bind to the insulin receptor. When the expression vectors ptkNEO and pAL-8 which incorporate the neo gene were complexed to the conjugate in an in vitro system of transfection, G418 resistant clones developed at frequencies of 2.0–5.5 × 10 −5 . Subsequently, a 923bp PstI fragment within the neo sequence was identified by Southern transfer in genomic DNA from transfected cell populations which had been maintained on a G418 regime for 44 days. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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