Skimmed milk as an alternative for IPTG in induction of recombinant protein expression
| Autor: | Mohammad-Hosein Khani, Masoumeh Bagheri |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Isopropyl Thiogalactoside
Salmonella typhimurium 0106 biological sciences food.ingredient Recombinant Fusion Proteins Genetic Vectors Gene Expression lac operon Lactose medicine.disease_cause 01 natural sciences Protein expression law.invention 03 medical and health sciences fluids and secretions food Bacterial Proteins law 010608 biotechnology Skimmed milk Protein purification Escherichia coli medicine Animals Histidine Cloning Molecular 030304 developmental biology 0303 health sciences biology Chemistry food and beverages biology.organism_classification Milk Lac Operon Biochemistry Toxicity Recombinant DNA bacteria Oligopeptides Bacteria Flagellin Biotechnology |
| Zdroj: | Protein Expression and Purification. 170:105593 |
| ISSN: | 1046-5928 |
| DOI: | 10.1016/j.pep.2020.105593 |
| Popis: | Cost-effectiveness is an important issue in biotechnological manufacturing industry and using alternative cheap materials with the same benefits has been noticed in most literatures. Isopropyl β- d -1-thiogalactopyranoside (IPTG), a well-known chemical element for induction of protein expression, has several disadvantages such as high expense and toxicity. In this study, we aimed to introduce skimmed milk as an alternative material for protein expression by induction of lac operon. In this way, Escherichia coli BL21 (DE3) bacteria were induced using 1 mM IPTG or 1.0% (w/v) skimmed milk. Protein purification was performed using Ni-NTA (nickel-nitrilotriacetic acid) for His-tagged recombinant proteins and protein purity was evaluated by SDS-PAGE. Results showed high level of recombinant protein expression using skimmed milk, and interestingly, the growth rate of bacteria improved. Our findings suggested that skimmed milk can be a suitable alternative for induction of recombinant protein expression, which has advantages such as more availability and affordability, in comparison to IPTG supplementation. |
| Databáze: | OpenAIRE |
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