摘要
差速离心和蔗糖密度梯度离心纯化经鸡胚成纤维细胞蚀斑克隆后的犬瘟热毒Ｓ＿８毒株，采用ＳＤＳ－ＰＡＥＧ电泳及Ｗｅｓｔｅｒｎｂｌｏｔ转印技术详细分析了犬瘟热病毒的结构多肽。回收凝胶中１条６３Ｋｄ的蛋白带，经３种不同的蛋白酶消化后，首次证明在犬瘟热病结构多肽中，融合蛋白Ｆ多肽的两个亚单位（Ｆ＿１和Ｆ＿２）是以胰蛋白酶的特异性水解位点精氨酸为桥梁连接形成Ｆ多肽的。试验结果表明，犬瘟热病毒主要包括Ｌ（２００．５Ｋｄ）、ＨＡ（７４Ｋｄ）、Ｐ（６８Ｋｄ）、Ｆ（６３Ｋｄ）、ＮＰ（５６Ｋｄ）、Ｍ（３５Ｋｄ）等６条结构多肽，同时下蛋白可在电泳中裂解成Ｆ＿１（４１Ｋｄ）和Ｆ＿２（２７Ｋｄ）２个亚单位。试验还发现其它３条分子量分别为５２Ｋｄ、４４Ｋｄ、３２Ｋｄ的病毒蛋白成分，它们是否均为病毒的结构多肽仍需深入研究。
The structural polypeptides of Canine Distemper virus (CDV)S_8 strain(derived from a Iivevaccine)were analysed by SDS-Polyacrylamide- Slab-gel electrophoresis and identified by Western-Blot withIabelled rabbits anti-CDV hyperimmune sera lgG as antibodies.One 63Kd polypeptides was obtained by elec-trotransfer and digested by trypsin,papain and pepsin to detect the subunits of CDV fusion protein. Test results demonstrate that CDV contains six major structural polypeptide as L. (200. 5kd)、 HA(74Kd)、P(68Kd)、F(63Kd)、NP(56Kd)、M(35Kd).F protein Contain F_1 (41Kd)and F_2(27Kd)two subunits. In addition,in the analysis,three other peptides with molecular weight as 52KD、44Dkand 32Kd were alsoi-dentified as CDV protein by western-blot.F_1adn F_2 subunits combinantion forms F protein on the site of Argand can be digested by trypsin.
出处
《中国兽医杂志》
CAS
北大核心
1995年第12期5-7,共3页
Chinese Journal of Veterinary Medicine
关键词
犬病
瘟病
病毒
结构多肽
蛋白亚单位
电泳
Canine Destemper Virus,structural polypeptides, protein subunits,SDS-polyacrylamide-Slab-gel eletrophoresis,Western-Blot.
