CAJ | 학술논문

以蓝绿温和胶电泳为工具,首次在国内用于条斑紫菜(Porphyra yezoensis)类囊体膜色素蛋白质复合物的研究。结果显示：(1)利用非离子型去污剂十二烷基麦芽糖苷(DM)增溶条斑紫菜类囊体膜, DM/Chla(w/w)15︰1产生的效果较好,在 BN-PAGE 胶中可以分离到较多较清晰的条带。(2)选取蔗糖密度层50%条带制备得到的类囊体膜样品进行第一向BN-PAGE和第二向SDS-Urea-PAGE电泳实验。第一向电泳分离出4个蛋白复合物,对第二向电泳胶上的15个蛋白点切取做质谱鉴定,检测到 PSⅡ47ku, PSⅡ44ku, cytochrome f , PSⅡ D2, PSⅡ D1等蛋白。本实验证实了温和胶电泳与SDS电泳结合对于条斑紫菜这种原始红藻的类囊体膜研究方面应用的可行性。
이람록온화효전영위공구,수차재국내용우조반자채(Porphyra yezoensis)류낭체막색소단백질복합물적연구。결과현시：(1)이용비리자형거오제십이완기맥아당감(DM)증용조반자채류낭체막, DM/Chla(w/w)15︰1산생적효과교호,재 BN-PAGE 효중가이분리도교다교청석적조대。(2)선취자당밀도층50%조대제비득도적류낭체막양품진행제일향BN-PAGE화제이향SDS-Urea-PAGE전영실험。제일향전영분리출4개단백복합물,대제이향전영효상적15개단백점절취주질보감정,검측도 PSⅡ47ku, PSⅡ44ku, cytochrome f , PSⅡ D2, PSⅡ D1등단백。본실험증실료온화효전영여SDS전영결합대우조반자채저충원시홍조적류낭체막연구방면응용적가행성。
The blue-native polyacrylamide gel electrophoresis was used for the first time in investigation of thylakoid membrane proteome of Porphyra yezoensis. The results showed that:(1) dodecyl-β-maltoside was a suitable detergent for the solubilization and stabilization of super-complexes of thylakoid membrane proteins. The detergent/Chla ratio of 15︰1 gave the best solubilizing effect and a series of clear protein bands were ob-tained using BN-PAGE assay;(2) The sample collected at 50%sucrose was separated by BN-PAGE in the first dimension and then SDS-Urea-PAGE in the second dimension. Four protein complexes were separated by 1-D BN-PAGE gels. 15 protein dots separated by 2-D SDS-Urea-PAGE were analyzed by MALDI-TOF MS and a series of proteins including PSⅡ, 47ku, PSⅡ, 44ku, cytochrome f, PSⅡ D2, PSⅡ and D1 were confirmed. These data demonstrated that combination of BN-PAGE with SDS-Urea-PAGE can be well used in study of thylakoid membrane proteome of P. yezoensis.