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《Acta Biologiae Experimentalis Sinica》 2001-01
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IMMUNOGOLD LOCALIZATION OF RIBULOSE-1,5-BISPHOSPHATE CARBORYLSAE/OXYGENASE IN CHLOROPLASTS OF CHLORELLA

HE Pei Min ZHANG Rong Xian ZHANG Da Bing ZHAO Jjian Hua LIANG Wuan Qi (Agriculture University of Nanjing, Nanjing 210095; Shanghai Fisheries University, Shanghai 200090; Agriculture Institute of Shanghai, Shanghai 200106)  
Ribulose 1, 5-bisphosphate carboxylase/oxygenase (Rubisco)is a first key enzyme in the Calvin Circle of plant cell photosynthesis. This paper mainly studied gold immunolocalizaion of Rubisco of Chlorella spp. 640909, and the Native-PAGE and,SDS-PAGE and Western blotting analysis, as well as the observation to pyrenoid ultra structure. The Native-PAGE result showed a main band, evidenced as the Rubisco band by the Western blot with the antibody against the Rubisco from C. prototothe-coides , The special immunoacton of Rubisco from Chlorella spp. 640909 and the antibody to large subunit of Rubisco from C. prothecoides showed the large subunit proteins of Rubisco in the two species of Chlorella shared the high homology. The SDS-PAGE and Western blotting maps showed the molecule weight of the large subunit of Rubisco of Chlorella spp. 640909 was about 55KD. The shape of pyrenoid ultra structure of the electronic microscope was oblong, and was embedded in starch sheath, with 2 swelling thylakoids through out a center portrait channel of the pyrenoid. There were some connections between pyrenoid and the chloroplast stroma. The distribution of the large subunits and the whole Rubisco in the chloroplast of Chrolella spp. 640909 was studied by immunoelectron microscopy by embedded sections with antibody to large subunit and whole enzyme followed by second antibody, goad anti-rabbit immunoglobulin G conjugated to 10nm gold particles(Sigma production). The result showed the antibodies against large subunit and whole enzyme heavily labeled the pyrenoid, as well as starch sheath region, whereas the thylakoid region of the plastid was lightly labeled. And the whole Rubisco antibody labeled the pyrenoid surface more heavily than the large subunit antibody did. It is demonstrated the pyrenoid and starch sheath have the photosynthesis function. Rubisco concentrating in pyrenoid and starch sheath is valuable to fix CO2 for photosynthesis in algae.
【Fund】: 上海市曙光计划(编号:SG-98-032);; 国家重点基础研究发展规划项目(编号:G1998010100)~~
【CateGory Index】: Q946
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