Difference between revisions of "Applications of Ribozymes in Synthetic Systems - Danielle Jordan"
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== How do you make artificial ribozymes? == | == How do you make artificial ribozymes? == | ||
The method of directed evolution is used to create specific ribozymes. Large quantities of RNA are produced using polymerase enzymes. The large library of ribozymes are mutated and amplified using error prone rtPCR (reverse transcriptase PCR). One method of selection is by using biotin tags, which are covalently bonded to a particular substrate and can then be extracted by streptavidin-magnetic beads. Thus, the molecules that exhibit the optimal ligase activity are recoved using the streptavidin matrix. | The method of directed evolution is used to create specific ribozymes. Large quantities of RNA are produced using polymerase enzymes. The large library of ribozymes are mutated and amplified using error prone rtPCR (reverse transcriptase PCR). One method of selection is by using biotin tags, which are covalently bonded to a particular substrate and can then be extracted by streptavidin-magnetic beads. Thus, the molecules that exhibit the optimal ligase activity are recoved using the streptavidin matrix. | ||
| + | == Why are they being used? == | ||
| + | Current protein promoters cannot easily be transferred from prokaryotic to eukaryotic organisms. However, ribozymes can be used in both systems because ribozymes do not rely on the cell's genetic information. Also, ribozymes can be artifically selected to respond to any set of exogenous molecules whereas there are only a limited number of protein promoters. | ||
| + | advantages | ||
| + | can be used in eukaryotic and prokaryotic cells | ||
| + | promoters can be efficiently selected for using directed evolution in response to a wide variety of exogenous molecules | ||
| + | do not have to be inserted into the DNA | ||
| + | difficulties | ||
| + | complex regulation pathway are difficult to construct | ||
| + | getting them into systems | ||
| + | not permanent | ||
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| + | <td>1</td> | ||
| + | <td>2</td> | ||
| + | </tr> | ||
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| + | <td>3</td> | ||
| + | <td>4</td> | ||
| + | </tr> | ||
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| + | <td>5</td> | ||
| + | <td>6</td> | ||
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| + | </table> | ||
Revision as of 05:21, 20 November 2007
Contents
What are ribozymes?
Ribozymes, also known as RNA enzymes or catalytic RNA, are RNA molecules that cataylze chemical reactions. They are able to catalzye hydrolysis of their own phosphodiester bonds or other RNA bonds. Some, such as RNA polymerase ribozymes, are able to catalyze their own synthesis.
History and Background
How do you make artificial ribozymes?
The method of directed evolution is used to create specific ribozymes. Large quantities of RNA are produced using polymerase enzymes. The large library of ribozymes are mutated and amplified using error prone rtPCR (reverse transcriptase PCR). One method of selection is by using biotin tags, which are covalently bonded to a particular substrate and can then be extracted by streptavidin-magnetic beads. Thus, the molecules that exhibit the optimal ligase activity are recoved using the streptavidin matrix.
Why are they being used?
Current protein promoters cannot easily be transferred from prokaryotic to eukaryotic organisms. However, ribozymes can be used in both systems because ribozymes do not rely on the cell's genetic information. Also, ribozymes can be artifically selected to respond to any set of exogenous molecules whereas there are only a limited number of protein promoters. advantages can be used in eukaryotic and prokaryotic cells promoters can be efficiently selected for using directed evolution in response to a wide variety of exogenous molecules do not have to be inserted into the DNA difficulties complex regulation pathway are difficult to construct getting them into systems not permanent
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