TRANSFERRING AND EXPRESSION OF GLUCOSE OXIDASE GENE ‐ GLUC IN PORPHYRA YEZOENSIS

Abstract

He, P.1, Yao, Q.2, Chen, Q.1, Guo, M., Xiong, A.2, Wu, W.1, Ma, J.1 1Shanghai Fisheries University, Shanghai 200090, PR China; 2Shanghai Agriculture Institute, Shanghai 201106, PR China; 3University of Connecticut, Connecticut 06901‐2315, USAThe disease‐resistant gene gluc, encoding glucose oxidase, has been used to increase advance crops' resistance to diseases. Now the gluc gene has been reconstructed in the plasmid pYP1203 between gus and cm+(cat) gene, a reporter gene and an antibiotics resistance labeling gene within the RB (right border) and LB (left border). With PEG and electroporation methods, the pYP1203 with gluc gene and gus, cat genes was transferred into the cells from thalli of Porphyra yezoensis. After 24‐72 hours, the GUS detection in transgenic cells of Porphyra yezoensis was positive. Cell transformation efficiency could reach as high as about 3‐4%. The expression of gus gene in the filaments regenerated from transgenic cells was also detected. Furthermore with the chloromycetin resistance screening, the transgenic cells of Porphyra survived over one month in the 50 ug/ml of chloromycetin concentration, while the wild cells could not survive in the same chloromycetin concentration. That results indicated that the gluc gene might be integrated into the chromosomes of Porphyra yezoensis. The molecular test of these transgenic algae and their disease resistance analysis are continuing.