Team:Penn State/MSC Design

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Revision as of 03:58, 4 October 2012

Multiple Start Codons Overview

Multiple Start Codons

A frustrating yet commonly observed phenomenon in the lab is the production of unexpected proteins. These occurrences may be explainable by multiple start codons in the mRNA strand. Codon slippage is a theory practically untouched by research, and this project aspires to shed some light on the issue.

Multiple Start Codons

Sample navigation menu:

Overview | Design | Results

Initial Construct

The initial construct for this project was designed using a basic vector and a relatively new product available from IDT (Integrated DNA Technologies), called gBlocks. gBlocks are synthesized dsDNA oligos with adjacent 40 base-pair overlap sequences. These oligos are particularly useful in multi-part constructions within a CBAR or Gibson Assembly reaction protocol.

The construct itself includes a promoter, ribosome binding site, and

Assembly

The oligos were ordered based on a digital representation of the desired plasmid sequence. The DNA sequence was manipulated through the free software ApE (A plasmid Editor) available through the University of Utah

Initial construction proved difficult. Whether this was due to inexperience or poor design, we cannot be sure. The final construct displaying a correct sequence has been assembled, however, and is giving us new insight into the codon slippage pheomenon.