Protein synthesis monitoring (psm)
First Claim
1. A method for monitoring protein synthesis in a protein synthesis system, the method comprising:
- providing a marker for protein synthesis in the system, said marker being detectable through detection of electromagnetic radiation;
detecting electromagnetic radiation emitted from the system; and
analyzing said emitted radiation to monitor protein synthesis activity in said system.
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Accused Products
Abstract
A method and a device are disclosed for monitoring the synthesis of proteins by the ribosome in real time, in vivo as well as in in-vitro. The ribosome is engineered to carry a donor fluorophore, and tRNA and/or amino acids and/or some other part of the ribosome are either engineered to carry acceptor fluorophores or else their natural fluorescent properties are utilized as acceptors. As the ribosomes mechanism processed the mRNA and tRNA molecules and synthesizes a polypeptide chain, a light source illuminates the ribosome, exciting the donor fluorophores and thereby the acceptor fluorophores whenever these are in sufficient proximity to a donor. The resulting signals are detected and used as a key for real-time database searching and identification of the protein being synthesized. The resulting data can be tabulated and interpreted in different ways. FIG. (1) describes the properties of a FRET pair and the dependence of FRET on pair distance.
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Citations
86 Claims
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1. A method for monitoring protein synthesis in a protein synthesis system, the method comprising:
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providing a marker for protein synthesis in the system, said marker being detectable through detection of electromagnetic radiation;
detecting electromagnetic radiation emitted from the system; and
analyzing said emitted radiation to monitor protein synthesis activity in said system. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 23, 24, 25, 26, 27, 28, 29, 30, 31, 62, 64)
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32. An apparatus for measuring protein synthesis by a protein synthesis system, said apparatus comprising:
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a container for containing a plurality of components for the system, wherein at least one component is capable of emitting electromagnetic radiation due to a protein synthesis activity;
a detection system to measure emitted radiation from the system; and
a computational device to analyze said emitted radiation and determine the protein synthesis activity in said system. - View Dependent Claims (33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 63)
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65. A method for ribosome labeling to allow protein synthesis monitoring, said method comprising:
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selecting a fluorescent probe;
selecting a location on at least one of a ribosomal RNA or on a ribosomal protein according to at least one of a characteristic of said probe or a characteristic of at least one of said ribosomal RNA or said ribosomal protein; and
attaching said probe to said location. - View Dependent Claims (66)
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67. A method for protein production monitoring, said method comprising:
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selecting a protein synthesis system for PSM analysis;
selecting a fluorescent probe;
selecting a location on at least one of a ribosomal RNA or on a ribosomal protein according to at least one of a characteristic of said probe or a characteristic of at least one of said ribosomal RNA or said ribosomal protein;
attaching said probe to said location to perform PSM; and
analyzing signals from said probe to monitor the protein synthesis system.
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68. A method for detecting protein synthesis in a protein synthesis system, the method comprising:
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providing a marker for protein synthesis in the system, said marker having a label;
attaching said marker to at least one component of the system; and
detecting said label to determine protein synthesis activity in the system.
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69-85. -85. (canceled)
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86. (canceled)
Specification