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以原核表达的番茄SlMC7重组蛋白为抗原,制备SlMC7多克隆抗体,采用ELISA和蛋白质点杂的方法,研究了番茄II型metacaspase蛋白(SlMC7)的抗体效价和表达特性,并对SlMC7蛋白及基因在番茄果实发育过程中的表达差异进行了研究,以期为后续的SlMC7在番茄果实中功能性研究提供材料基础和参考依据。结果表明:SlMC7多克隆抗体效价在256 000以上,在1∶10 000的稀释度下,可识别低至1.6ng的纯蛋白。对各时期番茄果实Western-blot和Real time PCR结果表明,相对于绿熟期,SlMC7蛋白及基因在破色期果实中表达量显著上升,表明SlMC7与番茄果实成熟存在一定相关性。
The S1MC7 polyclonal antibody was prepared by using the prokaryotic expression vector S1MC7 of recombinant tomato as an antigen, and the antibody titer and expression characteristics of the tomato type II metacaspase protein (SlMC7) were studied by ELISA and dot blotting. The effects of SlMC7 protein and gene The differences in the expression of tomato fruit during fruit development were studied in order to provide the material basis and reference for the subsequent study of the function of SlMC7 in tomato fruit. The results showed that the titer of the SlMC7 polyclonal antibody was over 256 000, and the pure protein of 1.6ng could be identified at a dilution of 1: 10,000. The results of Western-blot and Real time PCR of tomato fruits at different stages showed that the expression of SlMC7 protein and gene in shattering fruits significantly increased compared with the green ripe stage, indicating that there is a certain correlation between SlMC7 and tomato fruit ripening.