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Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly
 

Generation of Plasmid Vectors Expressing FLAG-tagged Proteins Under the Regulation of Human Elongation Factor-1α Promoter Using Gibson Assembly

Article DOI: 10.3791/52235-v • 10:18 min • February 9th, 2015
February 9th, 2015 •
1, 1
1Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center

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Synthesis of custom plasmids is labor and time consuming. This protocol describes the use of Gibson assembly cloning to reduce the work and duration of custom DNA cloning procedure. The protocol described also produces reliable tagged protein constructs for mammalian expression at similar cost to the traditional cut-and-paste DNA cloning.

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Keywords: Gibson Assembly Plasmid Vector FLAG-tagged Proteins Human Elongation Factor-1α (hEF1α) Promoter Mouse Embryonic Stem Cells (mESCs) CstF-64 CstF-64 Mutant DNA Cloning PCR Bacterial Transformation Restriction Digestion DNA Sequencing
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