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Replacing Standard Reporters from Molecular Cloning Plasmids with Chromoproteins for Positive Clone Selection.

Molecules. 2018; 
Tafoya-RamírezMargarita Daniela,Padilla-VacaFelipe,Ramírez-Salda?aAna Patricia,Mora-Gardu?oJosué Daniel,Rangel-Serranoángeles,Vargas-MayaNaurú Idalia,Herrera-GutiérrezLuz Janeth,FrancoBern
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Codon Optimization The coding sequences for AmilCP was codon optimized (Integrated DNA Technologies Codon Optimization tool, http://www.idtdna.com/CodonOpt) for E. coli but also with a CAI suitable for other bacterial organisms, assessed with GenScript Rare Codon Analysis Tool (https://www.genscript.com/tools/rare-codon-analysis) Get A Quote

摘要

Cloning and expression plasmids are the workhorses of modern molecular biology. Despite the pathway paved by synthetic biology, laboratories around the globe still relay on standard cloning techniques using plasmids with reporter proteins for positive clone selection, such as β-galactosidase alpha peptide complementation for blue/white screening or , which encodes for a toxic DNA gyrase. These reporters, when interrupted, serve as a positive clone detection system. In the present report, we show that molecular cloning plasmids bearing the coding sequence for a 25.4 kDa protein, AmilCP, encoded by a 685 bp gene, that is well expressed in , render blue-purple colonies. Using this reporter pro... More

關鍵詞

AmilCP chromoprotein,clone selection,molecular cloning,plasmid,protein expres
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