國立高雄大學圖資館 |

Language: English

Back

Engineering and analysis of lucifera...

Ignowski, Jolene Marie.

Engineering and analysis of luciferases as quantitative reporter genes in live mammalian cells.

Record Type:	Electronic resources : Monograph/item
Title/Author:	Engineering and analysis of luciferases as quantitative reporter genes in live mammalian cells.
Author:	Ignowski, Jolene Marie.
Description:	125 p.
Notes:	Chair: David V. Schaffer.
Notes:	Source: Dissertation Abstracts International, Volume: 66-10, Section: B, page: 5544.
Contained By:	Dissertation Abstracts International66-10B.
Subject:	Engineering, Chemical.
Online resource:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3190828
ISBN:	9780542343094

Engineering and analysis of luciferases as quantitative reporter genes in live mammalian cells.
Ignowski, Jolene Marie.

Engineering and analysis of luciferases as quantitative reporter genes in live mammalian cells. - 125 p.

Chair: David V. Schaffer.

Thesis (Ph.D.)--University of California, Berkeley, 2005.

One of the limitations of using a light-based signal in vivo is the absorption of photons by tissue and hemoglobin. The lowest absorption coefficient for hemoglobin occurs in the red region of the visible spectrum, while Fluc has an emission maximum at 560 nm. A red-shifted Fluc would enhance the sensitivity of an in vivo assay and also create the possibility of using various colors to monitor multiple events. In Chapter 4, we describe the evolution, expression, and imaging of several red-emitting Flucs (lambdamax=600 nm) in living mammalian cells.

ISBN: 9780542343094Subjects--Topical Terms:

226989
Engineering, Chemical.

Engineering and analysis of luciferases as quantitative reporter genes in live mammalian cells.
LDR:03032nmm _2200289 _450 001 170886
005 20061228142312.5
008 090528s2005 eng d
020 $a 9780542343094
035 $a 00242916
040 $a UnM $c UnM
100 0 $a Ignowski, Jolene Marie. $3 244917
245 1 0 $a Engineering and analysis of luciferases as quantitative reporter genes in live mammalian cells.
300 $a 125 p.
500 $a Chair: David V. Schaffer.
500 $a Source: Dissertation Abstracts International, Volume: 66-10, Section: B, page: 5544.
502 $a Thesis (Ph.D.)--University of California, Berkeley, 2005.
520 # $a One of the limitations of using a light-based signal in vivo is the absorption of photons by tissue and hemoglobin. The lowest absorption coefficient for hemoglobin occurs in the red region of the visible spectrum, while Fluc has an emission maximum at 560 nm. A red-shifted Fluc would enhance the sensitivity of an in vivo assay and also create the possibility of using various colors to monitor multiple events. In Chapter 4, we describe the evolution, expression, and imaging of several red-emitting Flucs (lambdamax=600 nm) in living mammalian cells.
520 # $a Reporter genes are genetically encoded proteins that are used to indicate the transcriptional activity of a gene or promoter. One example of a reporter gene is the luciferase from the North American firefly, Photinus pyralis . Firefly luciferase (Fluc) catalyzes the reaction of luciferin with oxygen to produce a photon of light. Fluc has been used in a variety of applications including gene delivery, signal transduction, and tumor growth analysis. While luciferase is often utilized in mammalian cells, little is known about its kinetic activity inside live cells. In Chapter 2, we analyzed the kinetics of Fluc inside living cells and developed a model to quantitatively predict the bioluminescence signal. We found the KM to be 1 mM, a value much higher than the 10 muM reported in vitro, and found the half-life of the luciferase to be 2 hours.
520 # $a While most of the studies involving luciferase use it to generate information about populations, luciferase also has the ability to kinetically monitor individual cells using an ultra-sensitive digital camera. In Chapter 3, we design and test an ultra-sensitive microscopic imaging system and find we are able to quantitatively distinguish different behaviors of individual mammalian cells. Specifically, we monitored the degradation of the Fluc signal using and found a population half-life of 2.5 hours; however, individual cells exhibited a diversity of half-lives ranging between 2 and 7.5 hours.
590 $a School code: 0028.
650 # 0 $a Engineering, Chemical. $3 226989
650 # 0 $a Chemistry, Biochemistry. $3 226900
690 $a 0487
690 $a 0542
710 0 # $a University of California, Berkeley. $3 212474
773 0 # $g 66-10B. $t Dissertation Abstracts International
790 $a 0028
790 1 0 $a Schaffer, David V., $e advisor
791 $a Ph.D.
792 $a 2005
856 4 0 $u http://libsw.nuk.edu.tw:81/login?url=http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3190828 $z http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3190828