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Introduction
of GFP
General
Applications of GFP
Quantitative
monitoring of gene expression
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Quantitative
monitoring of gene expression
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Why quantitative? --bioprocess issues
While GFP
as a reporter gene has mostly been used for qualitative studies,
its great potential as a quantitative, real time and on-line indicator
for protein production has been realized in recent years. Albano
et al. first demonstrated that GFP fluorescence could be used to quantitatively
monitor GFP-fusion protein production in E. coli (Albano et al., 1996;
Albano et al., 1998). Other work using GFP for monitoring and controlling
in E. coli were also successful (Poppenborg et al., 1997; Delisa et al.,
1999; Chae et al., 2000). Other than E. coli, GFP has also been used to
monitor protein production in yeast (Li et al., 2000) and insect cells
(Cha et al., 1997). Meanwhile, various methods and devices for quantitative
measurements of GFP were developed (Randers-Eichhorn et al., 1997; Craig
et al., 1997; Korf et al., 1997; Endow and Piston, 1998 and Knight et al.,
1999).
References:
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Albano CR, Randers-Eichhorn L, Bentley WE, and Rao G 1998.
Green fluorescent protein as a real time quantitative reporter of heterologous
protein production. Biotechnol. Prog. 14: 351-354.
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Albano CR, Randers-Eichhorn L, Chang Q, Bentley WE and Rao
G 1996. Quantitative measurement of green fluorescent protein expression
and chromophore cyclization. Biotechnol. Tech. 10: 953-958.
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Cha HJ, Pham MQ, Rao G, Bentley WE 1997. Expression of green
fluorescent protein in insect larvae and its application for foreign protein
production. Biotechnol. Bioeng. 56: 239-247.
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Cha HJ, Wu CF, Valdes JJ, Rao G, Bentley WE 2000. Observations
of green fluorescent protein as a fusion partner in genetically engineered
Escherichia coli: Monitoring protein expression and solubility. Biotechnol.
Bioeng. 67:565-574
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Chalfie M, Kain S 1998. Green fluorescent protein: properties,
applications, and protocols. New York: Wiley-Liss.
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DeLisa MP, Li J, Rao G, Weigand WA and Bentley WE 1999. Monitoring
GFP-operon fusion protein expression during high cell density cultivation
of Escherichia coli using on-line optical sensor. Biotechnol. Bioeng.
65: 54-64.
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Poppenborg L, Frieh K, Flaschel E 1997. The green fluorescent
protein is a versatile reporter for bioprocess monitoring. J. Biotechnol.
58: 79-88.
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