NaCl 처리로 벼 유묘에서 유도된 단백질의 분리와 정제

Purification of A Novel Protein Induced by Salt Treatment in Rice Seedling

수도의 내염성 품종 안나프르나에 NaCl 50 mM을 처리하여 유도된 단백질의 분리정제를 수행하여 한 개의 새로운 단백질을 분리 정제하였다. 그 결과는 다음과 같다 1. 10일된 유묘에 50mM NaCl을 48시간 처리하여 유도된 단백질의 존재를 전기영동을 통하여 확인하였다. 2. FPLC를 이용한 DEAE와 phenyl column을 이용하여 순수한 정제가 가능하였고, 이 과정을 통하여 전기영동상에서 단일 밴드를 나타내는 하나의 단백질을 정제하였다. 3. 이 단백질의 분자량이 64,000이라는 것을 Se-phdex-G 100 column chromatography를 통하여 확인하였다. 4. 이 단백질에 대한 단일 항체를 조제하여 고역가 항체를 분비하는 hybridoma 세포주 3개를 작성하고 isotype등 특성을 조사하였다.

The monoclonal antibody against a novel protein, which was induced in the roots of rice seedlings treated with NaCl, was produced. Oryzea sativa L. cv. Annapruna grains were sown on clean sands and grown for 10days and then the seedlings were soaked in 50mM NaCl aqueous solution. In 48hrs of the NaCl treatment, the roots were collected and homogenated with liquid nitrogen and extraction buffer. The homogenate was centrifuged and to the supernatant 75％ ammonium sulfate was added to pre-cipitate proteins. From these proteins a novel protein was purified through DEAE-ion chromatography and FPLC(Phenyl column). This protein appeared as a single band in the native electrophoresis. Using this protein as antigen, monoclonal antibody was produced. Five cell lines that secreted antibodies specifically bound to this protein were constructed.