Optimization of an In Vitro TranscriptionTranslation System Based on Sulfolobus solfataricus Cell Lysate
Joint Authors
Lo Gullo, Giada
Mattossovich, Rosanna
Perugino, Giuseppe
La Teana, Anna
Benelli, Dario
Londei, Paola
Source
Issue
Vol. 2019, Issue 2019 (31 Dec. 2019), pp.1-10, 10 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2019-02-11
Country of Publication
Egypt
No. of Pages
10
Main Subjects
Abstract EN
A system is described which permits the efficient synthesis of proteins in vitro at high temperature.
It is based on the use of an unfractionated cell lysate (S30) from Sulfolobus solfataricus previously well characterized in our laboratory for translation of pretranscribed mRNAs, and now adapted to perform coupled transcription and translation.
The essential element in this expression system is a strong promoter derived from the S.
solfataricus 16S/23S rRNA-encoding gene, from which specific mRNAs may be transcribed with high efficiency.
The synthesis of two different proteins is reported, including the S.
solfataricus DNA-alkylguanine-DNA-alkyl-transferase protein (SsOGT), which is shown to be successfully labeled with appropriate fluorescent substrates and visualized in cell extracts.
The simplicity of the experimental procedure and specific activity of the proteins offer a number of possibilities for the study of structure-function relationships of proteins.
American Psychological Association (APA)
Lo Gullo, Giada& Mattossovich, Rosanna& Perugino, Giuseppe& La Teana, Anna& Londei, Paola& Benelli, Dario. 2019. Optimization of an In Vitro TranscriptionTranslation System Based on Sulfolobus solfataricus Cell Lysate. Archaea،Vol. 2019, no. 2019, pp.1-10.
https://search.emarefa.net/detail/BIM-1122252
Modern Language Association (MLA)
Lo Gullo, Giada…[et al.]. Optimization of an In Vitro TranscriptionTranslation System Based on Sulfolobus solfataricus Cell Lysate. Archaea No. 2019 (2019), pp.1-10.
https://search.emarefa.net/detail/BIM-1122252
American Medical Association (AMA)
Lo Gullo, Giada& Mattossovich, Rosanna& Perugino, Giuseppe& La Teana, Anna& Londei, Paola& Benelli, Dario. Optimization of an In Vitro TranscriptionTranslation System Based on Sulfolobus solfataricus Cell Lysate. Archaea. 2019. Vol. 2019, no. 2019, pp.1-10.
https://search.emarefa.net/detail/BIM-1122252
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1122252