摘要
采用凝胶柱层析、聚丙烯凝胶电泳、免疫印迹(Western blotting)和免疫扩散等方法对施氏鲟卵黄蛋白原(Vi-tellogenin,Vg)及其相关蛋白(Yolk protein,YP)进行了研究。结果表明,施氏鲟血清Vg是一种糖脂磷蛋白,其相对分子量为410kD,由分子量为205kD的两个同源亚基组成。Vg的3种相关蛋白YP1、YP2和YP3。其中YP1相对分子量为370kD,是一种糖脂磷蛋白,由相对分子量为97kD和33kD的两个亚基构成。YP2是一种相对分子量为144kD的磷脂蛋白,由相对分子量为94kD和45kD的2个亚基构成。YP3为相对分子量为66kD的磷蛋白,由相对分子量为30kD的同源亚基构成。
Sturgeon is an ancient family（Acipenseridae） of fishes close to the divergence fish that eventually evolved into terrestrial animals and those which evolved into modern teleost species.Amur sturgeon vitellogenin（Vg） and its corresponding yolk protin（YP） products,YP1,YP2 and YP3 were isolated from serum of female at vitellogenesis and eggs from ovulated amur sturgeon,respectively.Amur sturgeon Vg was purified from female serum by a combination of precipitation in DW and gel filtration on sephadex G-200.Vg was confirmed to be a lipoglycophosphoprotein by staining with Red oil,Molecular Proes＇ Pro-Q Emerald 300 Glycoprotein Gel stain and Pro-Q Diamond Phosphoprotein Gel Stain,which had an apparent molecular mass of 410kD and appeared as one major band corresponding to 205kD after native-PAGE and SDS-PAGE,and confirmed by western blotting.Three yolk proteins derived from vitellogenin（YP1,YP2 and YP3）.Apparent molecular weights of YP1,YP2 and YP3 were 370kD,144kD and 66kD,respectively.After SDS-PAGE,YP1 appeared as two bands of 97kD and 33kD,while YP2 were resolved as two bands of 94kD and 45kD under reducing conditions.The characters of Yolk protein 1 is similar to amur sturgeon Vg,it is a lipoglycophosphoprotein that can be stained by Red oil,Molecular Proes＇ Pro-Q Emerald 300 Glycoprotein Gel stain and Pro-Q Diamond Phosphoprotein Gel Stain,while Yolk protein 2 was confirmed to be a phospholipoprotein.Yolk protein 3 appeared as one band of 30kD,and it could be stained only by Pro-Q Diamond Phosphoprotein Gel Stain.Antiserum to YP2 and YP3 cross reacted with vitellogenin suggested that YP2 and YP3 were the cleaved products of vitellogenin,while the characters of purified yolk protein indicated that YP2 and YP3 were amur sturgeon Lipovitellin and Phosvitin like,respectively.There are eight subunits of vitellin of amur sturgeon appeared in reduced SDS-PAGE,as four subunits in unreduced SDS-PAGE.The results of double immunodiffusiong using anti-Vg showed that amur sturgeon Vg was composed of YP1 and YP2,and YP3 had less antigenicity against anti-Vg.
出处
《水生生物学报》
CAS
CSCD
北大核心
2008年第5期750-759,共10页
Acta Hydrobiologica Sinica
基金
国家"十五"科技攻关项目(2004BA526B0113)资助
关键词
施氏鲟
卵黄蛋白原
脂磷蛋白
高磷蛋白
Acipenser schrenckii
Vitellogenin
Lipovitellin
Phosvitin
