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Introduction of GFP

General Applications of GFP

Quantitative monitoring of gene expression

Reference & Links

         Quantitative monitoring of gene expression 
  • 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:
  • 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. 
  • 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. 
  • 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. 
  • 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 
  • Chalfie M, Kain S 1998. Green fluorescent protein: properties, applications, and protocols. New York: Wiley-Liss. 
  • 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. 
  • Poppenborg L, Frieh K, Flaschel E 1997. The green fluorescent protein is a versatile reporter for bioprocess monitoring. J. Biotechnol. 58: 79-88.