不同生态型芦苇Rubisco免疫学特性差异  

Differences of Rubisco Immunological Characteristics in Different Ecotypes of Reeds

作　　者：李卫霞[1] 王海波[1] 浦铜良[1] 

机构地区：[1]兰州大学生命科学学院

出　　处：《西北植物学报》2006年第9期1764-1769,共6页Acta Botanica Boreali-Occidentalia Sinica

基　　金：国家自然科学基金项目(30470163)

摘　　要：以河西走廊荒漠地区不同生态型芦苇为研究材料,提取并纯化得Rubisco蛋白,经SDS-PAGE凝胶电泳将Rubisco大、小亚基分离,用Rubisco全酶蛋白及其大、小亚基分别注射昆明系雄性小白鼠制备抗体,经Western-blotting鉴定结果表明:(1)水芦Rubisco全酶抗体可与水芦、沙芦及菠菜Rubisco大亚基发生反应,而与小亚基均未见显色反应,且水芦显色最深,沙芦略浅,菠菜最浅;(2)水芦、沙芦Rubisco大亚基抗体可与水芦、沙芦、菠菜大亚基发生抗原交叉反应,且均不与小亚基发生反应,并且其与菠菜Rubisco大亚基的反应程度明显低于水芦和沙芦;(3)用与Rubisco大亚基抗体同样的制备方法,均未检测到水芦、沙芦Rubisco小亚基抗体的产生;(4)菠菜Rubisco全酶抗体可与菠菜、水芦、沙芦、水稻Rubisco大亚基均发生抗原交叉反应,但仅与其自身小亚基反应,且与菠菜Rubisco大亚基显色反应最深,水稻略浅,沙芦、水芦稍有反应.由此说明,水芦、沙芦Rubisco全酶蛋白及其大亚基免疫学特性差异较小,而与双子叶植物菠菜相比差异较大;水芦、沙芦Rubisco蛋白免疫化学决定簇的差异主要决定于小亚基上,且其小亚基不具有抗原活性或抗原活性较弱.The Rubiscos(Ribulose-1,5-bisphosphate carboxylase/oxygenases) in different ecotypes of reeds growing in the desert region of Hexi Corridor were extracted and purified and their large and small subunit were separated by SDS-PAGE;and then Rubiscos holoenzymes and their large and small subunits were separated injected into male white mice of Kunming line and their antibodies were generated in the mice.Western-blotting analysis of the antibodies showed that(1) the antibodies of Rubisco holoenzyme in swamp reed could react with the large subunits of Rubiscos in swamp reed,dune reed and spinach,but showed no color reactions with the small subunits of them;the deepest reaction color occurred with the large subunit of Rubisco in swamp reed,medium reaction color took place with the large subunits of Rubisco in dune reed and the lightest reaction color occurred with the large subunit of Rubisco in spinach;(2) Antigen cross reactions took place between the antibodies of the large subunits of Rubiscos in swamp reed and dune reed,and the large subunits of Rubiscos in swamp reed,dune reed and spinach,but no antigen cross reactions occurred between the antibodies of the large subunits of Rubiscos in swamp reed and dune reed and the small subunits of Rubiscos in swamp reed,dune reed and spinach;the antigen cross reaction with the large subunits of Rubiscos in spinach was clearly weaker than those in swamp reed and dune reed;(3) Although the same method as was used to generate the antibodies to the large subunits of Rubiscos was adopted to prepare the small subunits of Rubiscos,no antibodies to the small subunits of Rubiscos in swamp reed and dune reed were not detected;(4) Antigen cross reaction could occur between the antibody to Rubiscos holoenzyme in spinach and the large subunits of Rubiscos in spinach,dune reed,swamp reed and rice as well as the antibody to the small subunits of Rubisco in spinach;the deepest color occurred in the reaction with the large subunit of Rubisco in spinach,less deep color occurred in the reactio

关 键 词：RUBISCO Western-blotting 生态型 抗原活性 

分 类 号：Q948.1[生物学—植物学] S432.2[农业科学—植物病理学]