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Engineering Of Living Cells For The Expression Of Holo-phycobiliprotein-based Constructs

技术优势: Furthermore, these probes can be used in conjunction with GFPs or phytofluors [J.T. Murphy and J.C. Lagarias (1997) Curr. Biol. 7, 870-876; G. A. Gambetta and J. C.Lagarias (2001) Proc. Natl. Acad. Sci. USA, Vol. 98, 10566-10571] where it is desired to use multiple intracellular fluorescent labels, or in applications that require fluorescence energy transfer (FRET).

技术应用: Fluorescent protein probes expressed in vivo, such as Aequorea victoria green fluorescent protein (GFP) and its variants are of great value in many fields of research in cell biology, such as studies of protein localization in cells, cell division, organelle dynamics and vesicle trafficking, developmental biology, reporting gene expression, assessing uptake and expression of DNA in transfection experiments, etc.Phycobiliproteins can be used in all of the applications that currently use GFPs. Moreover, phycobiliproteins are superior fluorophores to GFPs. They absorb and emit at higher wavelengths than GFPs, and have comparable quantum yields but much higher extinction coefficients [see, A. J. Tooley, Y. A. Cai, and A. N. Glazer (2001) Proc. Natl. Acad. Sci. USA, Vol. 98, 10560-10565].

*Abstract: Purified native phycobiliproteins and their subunits fluoresce strongly and, since 1982, have been widely used as labels for cell sorting and analysis, and in a wide range of other fluorescence based assays. This invention embodies the use of phycobiliproteins and phycobiliprotein fusions as in vivo fluorescent protein probes.

*Principal Investigation: Name: Yuping Cai
Department:

Name: Alexander Glazer
Department:

Name: Aaron Tooley
Department:

附加资料: Patent Number: US6740507B2
Application Number: US2001919486A
Inventor: Glazer, Alexander N. | Tooley, Aaron J. | Cai, Yuping
Priority Date: 31 Jul 2001
Priority Number: US6740507B2
Application Date: 31 Jul 2001
Publication Date: 25 May 2004
IPC Current: C07K0014435 | C12N000902 | C12N000988 | C12N001552 | C12N001562 | C12P002104
US Class: 4350701 | 43500611 | 43500792 | 4352522 | 4352523 | 4353201 | 435325 | 435419 | 435449 | 435455 | 435468 | 435006
Assignee Applicant: The Regents of the University of California
Title: Engineering of living cells for the expression of holo-phycobiliprotein-based constructs
Usefulness: Engineering of living cells for the expression of holo-phycobiliprotein-based constructs
Summary: The cells are useful for expressing holo-phycobiliprotein-based constructs, useful in enzymology and chemistry of phycobiliprotein synthesis. The phycobiliproteins are useful as in vivo fluorescent protein probes.
Novelty: New recombinant cell comprising a heterologous-to-the-cell, fluorescent, first holo-phycobiliprotein domain fused a heterologous protein domain, useful for expressing a holo-phycobiliprotein fusion protein

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