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

杜宜殷(Yi-Yin Do) ； 黃鵬林(Pung-Ling Huang)

《中國園藝》 42卷2期 (1996/06) Pp. 162-174

https://doi.org/10.6964/JCSHS.199606.0162

摘要

構築蝴蝶之一互補DNA pOACO31於表達載體pET14b(+)中成爲嵌合基因(chimeric gene)，利用噬菌體(bacteriophage)T7啓動子(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。

關鍵字

原核表達系統 ； pET表達載體 ； 蛋白質純化

並列摘要

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.

並列關鍵字

Prokaryotic expression system ； pET expression vector ； Protein purification