Quantitative Screening of Insect Cell Transform ants Stably Expressing GFP_(uv)-β1,3-N-acetylglucosaminyltransferase 2 Fusion Protein

摘要

Insect cell transformants, stably expressing human β1,3-N-acetylglucosaminyltransferase 2 (β3GnT2) 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 β3GnT 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.