Biotechnology and Bioprocess Engineering:BBE

  • 제10권3호
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  • Pages.275-279
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  • 2005
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  • 1226-8372(pISSN)
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  • 1976-3816(eISSN)
한국생물공학회 (The Korean Society for Biotechnology and Bioengineering)
권호 표지

Quantitative Screening of Insect Cell Transformants Stably Expressing $GFP_{uv}-{\beta}1$, 3-N-acetylglucosaminyltransferase 2 Fusion Protein

  • Deo Vipin Kumar (Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University) ;
  • Kato Tatsuya (Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University) ;
  • Asari Naoko (Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University) ;
  • Park Enoch Y. (Laboratory of Biotechnology, Department of Applied Biological Chemistry, Faculty of Agriculture, Shizuoka University)
  • 발행 : 2005.06.01
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초록

Insect cell transformants, stably expressing human $GFP_{uv}-{\beta}1$, 3-N-acetylglucosaminyltransferase 2 $({\beta}3GnT2)$ as the green fluorescent protein $(GFP_{uv})-fused$ protein, were efficiently isolated on Western blot by the quantification of the densitometric intensity of the fusion protein. From almost 150 transformants containing the fusion gene linked to three different types of signal sequence, two transformants, Tn-pXme4a and -pX28a, were successfully selected, showing 8.3 and 8.6 mU/mL ${\beta}3GnT$ activity, respectively. This method requires a screening time almost one-half that required in the isolation of stably transformed cells with high expression levels, and at the same time allows the handling a large number of transformants.

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