Difference between revisions of "GBlock Design for Casein and FAM20C Kinase Genes"
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=Codon Optimization, Restriction Analysis and Flanking Sequences for Casein and FAM20C Kinase Genes= | =Codon Optimization, Restriction Analysis and Flanking Sequences for Casein and FAM20C Kinase Genes= | ||
==Overview== | ==Overview== | ||
In this section, we describe using published amino acid sequences for our proteins of interest to design DNA sequences | In this section, we describe using published amino acid sequences for our proteins of interest to design their DNA sequences in a form useful for the project. | ||
The design process includes codon optimization for expression in baker's yeast (''Saccharomyces cerevisiae'',) restriction analysis to ensure our DNA sequences aren't cleaved by the restriction enzymes used in this project or other common enzymes, incorporation of the FAKS yeast alpha-factor secretion signal to direct our protein products out of yeast cells, addition of restriction sites for Electra cloning, addition of BioBrick flanking sequences to allow assembly standardization, and disruption of Kex protease amino acid recognition sites, as needed. | The design process includes codon optimization for expression in baker's yeast (''Saccharomyces cerevisiae'',) restriction analysis to ensure our DNA sequences aren't cleaved by the restriction enzymes used in this project or other common enzymes, incorporation of the FAKS yeast alpha-factor secretion signal to direct our protein products out of yeast cells, addition of restriction sites for Electra cloning, addition of BioBrick flanking sequences to allow assembly standardization, and disruption of Kex protease amino acid recognition sites, as needed. | ||
Revision as of 08:57, 29 September 2014
Codon Optimization, Restriction Analysis and Flanking Sequences for Casein and FAM20C Kinase Genes
Overview
In this section, we describe using published amino acid sequences for our proteins of interest to design their DNA sequences in a form useful for the project.
The design process includes codon optimization for expression in baker's yeast (Saccharomyces cerevisiae,) restriction analysis to ensure our DNA sequences aren't cleaved by the restriction enzymes used in this project or other common enzymes, incorporation of the FAKS yeast alpha-factor secretion signal to direct our protein products out of yeast cells, addition of restriction sites for Electra cloning, addition of BioBrick flanking sequences to allow assembly standardization, and disruption of Kex protease amino acid recognition sites, as needed.
The final DNA sequences are submitted to Integrated DNA Technologies for synthesis as ["gBlocks Gene Fragments"]
A detailed description of the human and bovine casein and FAM20C kinase proteins with which we work can be found in Molecular biology.