원문정보
초록
영어
The (1,6)-fucosylation of core N-glycan (core fucose) has been known to play crucial roles in various pathophysiological events such as oncogenesis and metastasis. Therefore, a fluorescent probe recognizing core fucose will be a powerful tool for elucidating its functional role and further diagnosing cancer. The fleA gene, which has been known to code Aspergillus oryzae lectin (AOL), was cloned from A. oryzae RIB40. We fused it with the either citrine or cerulean gene (improved variants of yellow or cyan fluorescent protein) to generate two expression vectors. The resulting fusion gene products (FleA-citrin & FleA–cerulean) were successfully expressed in Escherichia coli and purified homogeneously using two step affinity purification method. The fluorescent FleA fusion proteins were shown to detect core fuose in glycoproteins on the surface of cancer cells. The reduction of core fucose amount induced by the transfection of (1,6)-fucosyltransferase gene, Fut8, siRNA was also successfully detected by the FleA fusion proteins. These results show that our FleA-fluorescent fusion proteins would be employedas a valuable fluorescent probe for detecting core fucose.