Production of red fluorescent protein (mCherry) in an inducible E. coli expression system in a bioreactor, purification and characterization
Yazarlar (5)
Tokat Gaziosmanpaşa Üniversitesi, Türkiye
Tokat Gaziosmanpaşa Üniversitesi, Türkiye
Bartın Üniversitesi, Türkiye
Tokat Gaziosmanpaşa Üniversitesi, Türkiye
Prof. Dr. İsa GÖKÇE
Tokat Gaziosmanpaşa Üniversitesi, Türkiye
| Makale Türü |
Özgün Makale
|
| Makale Alt Türü | Ulusal alan endekslerinde (TR Dizin, ULAKBİM) yayınlanan tam makale |
| Dergi Adı | International Advanced Researches and Engineering Journal |
| Dergi ISSN | 2618-575X |
| Dergi Tarandığı Indeksler | TR DİZİN |
| Makale Dili | İngilizce |
| Basım Tarihi | 04-2019 |
| Cilt No | 3 |
| Sayı | 1 |
| Sayfalar | 20 / 25 |
| Makale Linki | http://dergipark.gov.tr/download/article-file/684032 |
| Özet |
| New and improved genetic engineered variants of fluorescent proteins (FPs) have become usefultools for bioimaging in biomedical researches. Red fluorescent proteins (RFPs) first derivedfrom the sea anemone Discosoma show high performance in vivo labeling and imaging. mCherryis a member of RFPs which has very high photostability, resistant to photo bleaching and rapidmaturation. These advantages ensure that mCherry can be successfully fused to many proteinsand widely used for quantitative imaging techniques. In this study, the constructed recombinantplasmid pBADCherry was expressed in Escherichia coli BL21(AI) then culture conditions,inducer concentration and induction time were optimized. Results of sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) analysis demonstrated that 5 hours induction at0.04% of arabinose concentration was optimal for the highest mCherry yield. The expression ofhexa histidine-tagged (6xHis) recombinant mCherry was induced by arabinose and purificationperformed using nickel (Ni2+) affinity chromatography. High throughput expression of 81 mgfluorescent protein from a liter of E. coli culture carried out in bioreactor. |
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