Ge Healthcare UK Limited

Cancer Research Technology Limited

Ge Healthcare Limited

Amersham Plc

Anne Elizabeth Jones

Nicholas Thomas

Nigel Paul Michael

Simon Lawrence John Stubbs

Rahman Aziz Ismail

Ian D Goodyer

Michael John Francis

Jonathan Mark Kendall

Jonathon Noe Pines

Simon Lawrence Stubbs

The present invention provides novel engineered derivatives of green fluorescent protein (GFP) which have an amino acid sequence which is modified by amino acid substitution compared with the amino acid sequence of wild type Green Fluorescent Protein (SEQ ID NO: 2). The modified GFPs exhibit enhanced fluorescence relative to wtGFP when expressed in non-homologous cells at temperatures above 30&#xb0; C., and when excited at about 490 nm compared to the parent proteins, i.e. wtGFP. An example of a preferred protein is F64L-S175G-E222G-GFP. The modified GFPs provide a means for detecting GFP reporters in mammalian cells at lower levels of expression and/or increased sensitivity relative to wtGFP. This greatly improves the usefulness of fluorescent proteins in studying cellular functions in living cells.

Fluorescent proteins and methods of using same

The invention provides a novel, non-destructive and dynamic process for determining the cell cycle position of living cells. The invention also provides DNA constructs, and cell lines containing such constructs, that exhibit activation and deactivation of a detectable reporter molecule in a cell cycle specific manner. The invention thus allows greater precision in determining cell cycle phase status than existing techniques and further provides a method for continuous monitoring of cell cycle progression in individual cells.

Method for determining cell cycle position

The present invention provides novel engineered derivatives of green fluorescent protein (GFP) which have an amino acid sequence which is modified by amino acid substitution compared with the amino acid sequence of wild type Green Fluorescent Protein. The modified GFPs exhibit enhanced fluorescence relative to wtGFP when expressed in non-homologous cells at temperatures above 30&#xb0; C., and when excited at about 490 nm compared to the parent proteins, i.e. wtGFP. An example of a preferred protein is F64L-S175G-E222G-GFP. The modified GFPs provide a means for detecting GFP reporters in mammalian cells at lower levels of expression and/or increased sensitivity relative to wtGFP. This greatly improves the usefulness of fluorescent proteins in studying cellular functions in living cells.

Fluorescent proteins

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Amersham, United Kingdom

Anne Elizabeth Jones