AAS ›› 2015, Vol. 46 ›› Issue (2): 196-201.doi: 10.16098/j.issn.0529-1356.2015.02.009

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Analysis of expression of the different red fluorescent proteins in yeast cells

WU Qian-qian WANG Jing-song WANG Rui-kun LU Yi-xin LI Xiang-ming*   

  1. Department of Preventive Medicine,Medical College of Yangzhou University, Jangsu Yangzhou 225001,China
  • Received:2014-03-24 Revised:2014-11-05 Online:2015-04-06 Published:2015-04-06
  • Contact: LI Xiang-ming E-mail:yzzpt@sina.com

Abstract:

Objective To compare the expression of fluorescent protein gene in yeast cells. Methods Four of red fluorescent protein (DsRed) yeast expression vectors, pGPD-DsRed, pGPD-DsRed-express-2, pGPD-yDsred and pGPD-yDsred-express-2, were constructed using the molecular biology method. pGPD-yDsRed and pGPD-yDsRed-express-2 contained the yeast cell preference codon. These 4 vectors were transformed into W303-1A yeast cells and DsRed expression was observed under the fluorescence microscope. Multifunctional microplate reader was used to measure the luminous intensity of yeast cells. Results The fluorescence intensities of the 4 the DsRed were significantly different. DsRed-express-2 emission was the strongest, followed by yDsRed-Express-2, and the weakest luminous intensity was yDsRed. Conclusion Red fluorescent protein strength has nothing to do with the codon preference in yeast cells. The best way is to select reporter gene of DsRed-express-2 to quantify the expression intensity of red fluorescent protein.

Key words: Red fluorescent protein, Codon preference, Yeast cells, RT-PCR