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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)
Publication Information
Biotechnology and Bioprocess Engineering:BBE / v.10, no.3, 2005 , pp. 275-279 More about this Journal
Abstract
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.
Keywords
3-N-acetylglucosaminyltransferase 2; insect cell; green fluorescent protein; screening; Western blot;
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