Title

Why is it so dirty? Purification of Proteins

Poster Number

14

Format

Poster Presentation

Abstract/Artist Statement

The underlying theme of biotechnology is the understanding of cellular functions and ways to manipulate the mechanics to produce pragmatic products. In order to study these mechanics, which are produced naturally by living cells and microbes, it is necessary to replicate or amplify the apparatuses that yield these products. In general, these apparatuses are usually proteins and enzymes. Through various amplification and purification steps, it is possible to obtain a very concentrated amount of proteins that can further the investigation of various cellular functions. First, the organism used is typically the bacterial strain Escherichia coli used to carry the plasmid encoding for the gene to produce the target protein. Depending on the type of promoter located on the plasmid, expression of the plasmid can be induced by certain natural biochemicals. IPTG is generally used as the induction factor, causing the E.coli cells to yield the wanted protein in large quantities. Through experimentation, different laboratory techniques were utilized to purify the protein. Ideally, the protein would have been put through a Q-sepharose fast flow column, followed by dialysis, and finally purified using a BioRex 70 column in order to remove cellular debris and impurities. To ensure the protein was present, PCR and agarose gel electrophoresis was performed during the intermediate purification processes. It was discovered that the protein achieved optimal activity through dialysis and alternative methods without utilizing either the Q- sepharose fast flow column or BioRex 70 column.

Location

Pacific Geosciences Center

Start Date

5-5-2007 1:00 PM

End Date

5-5-2007 3:00 PM

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May 5th, 1:00 PM May 5th, 3:00 PM

Why is it so dirty? Purification of Proteins

Pacific Geosciences Center

The underlying theme of biotechnology is the understanding of cellular functions and ways to manipulate the mechanics to produce pragmatic products. In order to study these mechanics, which are produced naturally by living cells and microbes, it is necessary to replicate or amplify the apparatuses that yield these products. In general, these apparatuses are usually proteins and enzymes. Through various amplification and purification steps, it is possible to obtain a very concentrated amount of proteins that can further the investigation of various cellular functions. First, the organism used is typically the bacterial strain Escherichia coli used to carry the plasmid encoding for the gene to produce the target protein. Depending on the type of promoter located on the plasmid, expression of the plasmid can be induced by certain natural biochemicals. IPTG is generally used as the induction factor, causing the E.coli cells to yield the wanted protein in large quantities. Through experimentation, different laboratory techniques were utilized to purify the protein. Ideally, the protein would have been put through a Q-sepharose fast flow column, followed by dialysis, and finally purified using a BioRex 70 column in order to remove cellular debris and impurities. To ensure the protein was present, PCR and agarose gel electrophoresis was performed during the intermediate purification processes. It was discovered that the protein achieved optimal activity through dialysis and alternative methods without utilizing either the Q- sepharose fast flow column or BioRex 70 column.