Fluorescence Assay for Polymerase Arrival Rates
To engineer complex synthetic biological systems will require modular design, assembly, and characterization strategies. The RNA polymerase arrival rate (PAR) is defined to be the rate that RNA polymerases arrive at a specified location on the DNA. Designing and characterizing biological modules in...
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| Language: | en_US |
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2004
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| Online Access: | http://hdl.handle.net/1721.1/7112 |
| _version_ | 1811093356149735424 |
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| author | Che, Austin |
| author_facet | Che, Austin |
| author_sort | Che, Austin |
| collection | MIT |
| description | To engineer complex synthetic biological systems will require modular design, assembly, and characterization strategies. The RNA polymerase arrival rate (PAR) is defined to be the rate that RNA polymerases arrive at a specified location on the DNA. Designing and characterizing biological modules in terms of RNA polymerase arrival rates provides for many advantages in the construction and modeling of biological systems. PARMESAN is an in vitro method for measuring polymerase arrival rates using pyrrolo-dC, a fluorescent DNA base that can substitute for cytosine. Pyrrolo-dC shows a detectable fluorescence difference when in single-stranded versus double-stranded DNA. During transcription, RNA polymerase separates the two strands of DNA, leading to a change in the fluorescence of pyrrolo-dC. By incorporating pyrrolo-dC at specific locations in the DNA, fluorescence changes can be taken as a direct measurement of the polymerase arrival rate. |
| first_indexed | 2024-09-23T15:43:57Z |
| id | mit-1721.1/7112 |
| institution | Massachusetts Institute of Technology |
| language | en_US |
| last_indexed | 2024-09-23T15:43:57Z |
| publishDate | 2004 |
| record_format | dspace |
| spelling | mit-1721.1/71122019-04-12T08:33:59Z Fluorescence Assay for Polymerase Arrival Rates Che, Austin AI To engineer complex synthetic biological systems will require modular design, assembly, and characterization strategies. The RNA polymerase arrival rate (PAR) is defined to be the rate that RNA polymerases arrive at a specified location on the DNA. Designing and characterizing biological modules in terms of RNA polymerase arrival rates provides for many advantages in the construction and modeling of biological systems. PARMESAN is an in vitro method for measuring polymerase arrival rates using pyrrolo-dC, a fluorescent DNA base that can substitute for cytosine. Pyrrolo-dC shows a detectable fluorescence difference when in single-stranded versus double-stranded DNA. During transcription, RNA polymerase separates the two strands of DNA, leading to a change in the fluorescence of pyrrolo-dC. By incorporating pyrrolo-dC at specific locations in the DNA, fluorescence changes can be taken as a direct measurement of the polymerase arrival rate. 2004-10-20T20:32:21Z 2004-10-20T20:32:21Z 2003-08-31 AITR-2003-017 http://hdl.handle.net/1721.1/7112 en_US AITR-2003-017 112 p. 92476964 bytes 3362118 bytes application/postscript application/pdf application/postscript application/pdf |
| spellingShingle | AI Che, Austin Fluorescence Assay for Polymerase Arrival Rates |
| title | Fluorescence Assay for Polymerase Arrival Rates |
| title_full | Fluorescence Assay for Polymerase Arrival Rates |
| title_fullStr | Fluorescence Assay for Polymerase Arrival Rates |
| title_full_unstemmed | Fluorescence Assay for Polymerase Arrival Rates |
| title_short | Fluorescence Assay for Polymerase Arrival Rates |
| title_sort | fluorescence assay for polymerase arrival rates |
| topic | AI |
| url | http://hdl.handle.net/1721.1/7112 |
| work_keys_str_mv | AT cheaustin fluorescenceassayforpolymerasearrivalrates |