Flow cytometric analysis of human lysozyme production in recombinantSaccharomyces cerevisiae |
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Authors: | Marvin S. Peterson Myoung-Dong Kim Ki-Cheol Han Ji-Hyun Kim Jin-Ho Seo |
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Affiliation: | (1) Department of Food Science and Technology and Research Center for New Bio-Materials in Agriculture, Seoul National University, 441-744 Suwon, Korea;(2) Signal Pharmaceuticals Inc., 92121 San Diego, CA, USA;(3) Ministry of Commerce, Industry and Energy, 427-010 Kwacheon, Korea |
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Abstract: | Flow cytometric techniques were used to investigate cell size, protein content and cell cycle behavior of recombinantSaccharomyces cerevisiae strains producing human lysozyme (HLZ). Two different signal sequences, the native yeastMFα1 signal sequence and the rat α-amylase signal sequence, were used for secretion of HLZ. The strain containing the rat α-amylase signal sequence showed a higher level of internal lysozyme and lower specific growth rates. Flow cytometric analysis of the total protein content and cell size showed the strain harboring the native yeast signal sequence had a higher total protein content than the strain containing the rat α-amylase signal sequence. Cell cycle analysis indicated that the two lysozyme producing recombinant strains had an increased number of cells in the G2+M phase of the yeast cell cycle compared with the host strain SEY2102. |
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Keywords: | flow cytometry Saccharomyces cerevisiae human lysozyme cell cycle secretion |
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