کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
2824044 1570337 2016 9 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
pNEB193-derived suicide plasmids for gene deletion and protein expression in the methane-producing archaeon, Methanosarcina acetivorans
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی ژنتیک
پیش نمایش صفحه اول مقاله
pNEB193-derived suicide plasmids for gene deletion and protein expression in the methane-producing archaeon, Methanosarcina acetivorans
چکیده انگلیسی


• We have created a suite of user-friendly plasmids for methanogens.
• The new plasmids are now compatible with ligation-independent cloning.
• We validated plasmids for markerless gene deletion.
• Plasmids were used to express a native and his-tagged reporter gene.

Gene deletion and protein expression are cornerstone procedures for studying metabolism in any organism, including methane-producing archaea (methanogens). Methanogens produce coenzymes and cofactors not found in most bacteria, therefore it is sometimes necessary to express and purify methanogen proteins from the natural host. Protein expression in the native organism is also useful when studying post-translational modifications and their effect on gene expression or enzyme activity. We have created several new suicide plasmids to complement existing genetic tools for use in the methanogen, Methanosarcina acetivorans. The new plasmids are derived from the commercially available Escherichia coli plasmid, pNEB193, and cannot replicate autonomously in methanogens. The designed plasmids facilitate markerless gene deletion, gene transcription, protein expression, and purification of proteins with cleavable affinity tags from the methanogen, M. acetivorans.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Plasmid - Volumes 84–85, March–May 2016, Pages 27–35
نویسندگان
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