摘要
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。
SlMC7 prokaryotic expression recombinant protein was used as test materials.SlMC7 polyclonal antiserum was obtained from immunized mice by using purified SlMC7 recombinant protein fromEscherichia coli.ELISA and protein dot plot methods were used to detect the antibody titer and expression characteristics and the expression difference at different stages,providing material foundation and reference basis for subsequent SlMC7 functional research in tomato fruits.The results showed that the titer of SlMC7 polyclonal antibodies was more than 1∶256 000and sensitivity of SlMC7 polyclonal antibodies was 1.6ng at the dilution of 1∶10 000.SlMC7 protein was detected at different stages,SlMC7 had a higher expression in breaker fruits.SlMC7 might be relevant to the fruit ripening of tomato.
出处
《北方园艺》
CAS
北大核心
2017年第16期50-55,共6页
Northern Horticulture
基金
国家自然科学基金资助项目(31672207)
