蝴蝶蘭OACO31嵌合基因之構築及抗體之製備

Construction of a Phalaenopsis OACO31 Chimeric Gene and Preparation of Antibody against OACO31

構築蝴蝶之一互補DNA pOACO31於表達載體pET14b（+）中成為嵌合基因（chimeric gene），利用噬菌體（bacteriophage）T7啟動子（promoter），於原核表達系統即大腸桿菌（Escherichia coli）中進行大量表現蛋白質，經電泳純化後，將所取得之60KD蛋白質注射至天竺鼠，製備成OACO31之抗體。

The coding region of a cDNA pOACO31 from Phalaenopsis was constructed into prokaryotic expression vector pET14b(+). After transformed into Escherichia coli, this chimeric gene was induced with isopropy-β-D-thiogalactopyranoside and the fusion protein exist as insoluble inclusion body. The 60KDa OACO31 polypeptide purified by Ni(superscript 2+) affinity cloumn and electrophoresis was used as an antigen to immunize guinea pig to obtain antibody for Western analysis. It will be useful for immunolocalization in tissues of the orchid.