殺蟲性 結晶蛋白質(Cry ⅡA) 特異的 抗體 生産 = Production of Antibodies against Insecticidal Crystal Paotein (Cry Ⅱ A)

다국어 초록 (Multilingual Abstract)

Bacillus thuringiensis, a gram-positive soil bacterium, is characterized by its ability to produce crystalline inclusions during sporulation. The crystal proteins exhibit a highly specific insecticidal activity. A crystal protein, Cry Ⅱ A, is specifically toxic to both lepidopteran and dipteran insects. Scientists are working hard to generate the insect-resistant transgenic plants due to the instability and low-persistence of the proteins. In this study, to confirm the expression of the cryⅡA gene, anti-CryⅡ A antibody was raised in a rabbit. The Cry Ⅱ A crystal protein was purified from E. coli JM103 harboring the cry ⅡA gene byc differential solubility. The Cry Ⅱ A was digested with trypsin for the activation. Some 300 ㎍ of the activated Cry Ⅱ A was mixed with the same amount of FCA, and the slurry was hyperdermally injected onto about 20 spots of the back, one spot of the thigh, and one spot of the footpad of the rabbit. The immunization was performed four times with two weeks interval. From the third immunization, FIC was substituted for the FCA. The rabbit serum was collected from the ear vein, and anti-Cry Ⅱ A antibody was purified by Protein A affinity chromatography. The anti-Cry Ⅱ A antibody recognized nanogram quantity (1.25 ng) of Cry Ⅱ A by solid phase immunoassay. So it can be available for screening of transgenic plants with the insecticidal characteristics of Cry Ⅱ A.

목차 (Table of Contents)

• I . 서 론
• Ⅱ. 재료 및 방법
• Ⅲ. 결 과
• Ⅳ. 고 찰