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Publikasi Staf Biotek

Expression and characterization of the Renilla luciferase with the cumulative mutation



Luciferase from Renilla reniformis (RLuc) is a good research tool as a reporter protein and bioimaging probes, yielding blue light using the substrate coelenterazine. However, the applications are limited since RLuc is unstable under various conditions. Therefore, an attempt was made to increase RLuc thermostability. In this study, 5 mutations reported previously [1] and one mutation obtained using site-directed mutagenesis were combined. As a result of this combination, the thermostability effect increased, with the mutant showing approximately 10°C higher stability. Furthermore, the mutant simultaneously improved a tolerance for protease digestion, e.g. trypsin and proteinase K, and for organic solvent. Residual activity of the mutant after treatment with 10 persen 2propanol, 10 persen DMF and 20 persen DMSO at 35°C for 1h was 29.4, 24.8 and 91.3 persen, respectively, whereas that of the wild type was 0.4, 0.1 and 24.3 persen, respectively.

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Judul Seri
Protein Expression and Purification
No. Panggil
-
Penerbit : .,
Deskripsi Fisik
145: 39-44.
Bahasa
English
ISBN/ISSN
-
Klasifikasi
NONE
Tipe Isi
text
Tipe Media
other
Tipe Pembawa
online resource
Edisi
-
Subyek
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-
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