摘要
Purification of photosystem Ⅱ （PSII） core complexes is a time-consuming and low-efficiency process. In order to isolate pure and active PSll core complexes in large amounts, we have developed a fast method to isolate highly active monomeric and dimeric PSll core complexes from spinach leaves by using sucrose gradient ultracentrifugation. By using a vertical rotor the process was completed significantly faster compared with a swing-out rotor. In order to keep the core complexes in high activity, the whole isolation procedure was performed in the presence of glycine betain and pH at 6.3. The isolated pigment-protein complexes were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, absorption spectroscopy, 77 K fluorescence spectroscopy and high performance liquid chromatography. Our results show that this method is a better choice for quick and efficient isolation of functionally active PSII core complexes.
Purification of photosystem Ⅱ （PSII） core complexes is a time-consuming and low-efficiency process. In order to isolate pure and active PSll core complexes in large amounts, we have developed a fast method to isolate highly active monomeric and dimeric PSll core complexes from spinach leaves by using sucrose gradient ultracentrifugation. By using a vertical rotor the process was completed significantly faster compared with a swing-out rotor. In order to keep the core complexes in high activity, the whole isolation procedure was performed in the presence of glycine betain and pH at 6.3. The isolated pigment-protein complexes were characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, absorption spectroscopy, 77 K fluorescence spectroscopy and high performance liquid chromatography. Our results show that this method is a better choice for quick and efficient isolation of functionally active PSII core complexes.
基金
supported by grant from the National Natural Science Foundation of China(30470149 and 30500101)
