دورية أكاديمية
أعرض في EDS

Construction and validation of the Tn5-P LtetO-1 -msfGFP transposon as a tool to probe protein expression and localization.

التفاصيل البيبلوغرافية
العنوان: Construction and validation of the Tn5-P LtetO-1 -msfGFP transposon as a tool to probe protein expression and localization.
المؤلفون: Passaris I; Laboratory of Food Microbiology, Department of Microbial and Molecular Systems, KU Leuven. Faculty of Bioscience Engineering, Kasteelpark Arenberg 22, 3000 Leuven, Belgium., Tadesse WM; Laboratory of Food Microbiology, Department of Microbial and Molecular Systems, KU Leuven. Faculty of Bioscience Engineering, Kasteelpark Arenberg 22, 3000 Leuven, Belgium., Gayán E; Laboratory of Food Microbiology, Department of Microbial and Molecular Systems, KU Leuven. Faculty of Bioscience Engineering, Kasteelpark Arenberg 22, 3000 Leuven, Belgium., Aertsen A; Laboratory of Food Microbiology, Department of Microbial and Molecular Systems, KU Leuven. Faculty of Bioscience Engineering, Kasteelpark Arenberg 22, 3000 Leuven, Belgium. Electronic address: abram.aertsen@kuleuven.be.
المصدر: Journal of microbiological methods [J Microbiol Methods] 2019 Jun; Vol. 161, pp. 56-62. Date of Electronic Publication: 2019 Apr 18.
نوع المنشور: Journal Article; Research Support, Non-U.S. Gov't
اللغة: English
بيانات الدورية: Publisher: Elsevier Biomedical Country of Publication: Netherlands NLM ID: 8306883 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1872-8359 (Electronic) Linking ISSN: 01677012 NLM ISO Abbreviation: J Microbiol Methods Subsets: MEDLINE
أسماء مطبوعة: Original Publication: Amsterdam, The Netherlands : Elsevier Biomedical, c1983-
مواضيع طبية MeSH: Bacterial Proteins/*genetics , Bacterial Proteins/*metabolism , DNA Transposable Elements/*genetics , Proteomics/*methods, Chromosome Mapping ; Conjugation, Genetic ; Genetic Markers ; Green Fluorescent Proteins/genetics ; Lac Operon ; Mutagenesis, Insertional ; Open Reading Frames ; Promoter Regions, Genetic
مستخلص: In this study we report the design, construction and validation of a novel transposon aimed to systematically screen for protein localization and expression patterns in prokaryotes using fluorescence microscopy. Upon random insertion in an open reading frame in the proper frame and orientation, the transposon creates an N-terminal fluorescent protein fusion to the msfGFP reporter. Moreover, in order to examine the localization of fusion proteins whose native expression might be too low or absent, the transposon was fitted with a P LtetO-1 promoter that makes the expression of the generated fluorescent protein fusions controllable by anhydrotetracycline. Importantly, upon flipping out the P LtetO-1 promoter and neighboring antibiotic resistance marker, an in-frame "sandwich" msfGFP fusion is created in which the N- and C-terminal portions of the targeted protein are again controlled by its native promoter.
(Copyright © 2019 Elsevier B.V. All rights reserved.)
فهرسة مساهمة: Keywords: Anhydrotetracycline inducible promoter; Cellular localization; Fluorescent protein fusion; Transposon mutagenesis
المشرفين على المادة: 0 (Bacterial Proteins)
0 (DNA Transposable Elements)
0 (Genetic Markers)
147336-22-9 (Green Fluorescent Proteins)
تواريخ الأحداث: Date Created: 20190421 Date Completed: 20200601 Latest Revision: 20200601
رمز التحديث: 20240628
DOI: 10.1016/j.mimet.2019.04.012
PMID: 31004623
قاعدة البيانات: MEDLINE
الوصف
تدمد:1872-8359
DOI:10.1016/j.mimet.2019.04.012