摘要
从我国分离到的一株单纯疱疹病毒Ⅰ型（ＨＳＶ－１－１６８株）病毒基因组中，分离出含有糖蛋白Ｄ（ｇＤ）基因的１．２ｋｂ片段，插入带有痘苗病毒天坛株ＴＫ区的质粒ｐＪＳＢ１１７５Ｐ７．５ｋ启动子下游，转染无白血病鸡胚原代细胞，获得带有ＨＳＶ－１－１６８ｇＤ基因的重组痘苗病毒。此株重组病毒在感染细胞膜上表达ＨＳＶ－１－１６８ｇＤ糖蛋白抗原，能与特异性单克隆抗体反应。在感染细胞中表达的膜抗原经ＳＤＳ－ＰＡＧＥ分析，表达分子量为５４ｋＤ糖蛋白。用Ｓｏｕｔｈｅｒｎ杂交分析了重组病毒ＤＮＡ中特异的ｇＤ基因，对作为活疫苗的重组痘苗病毒株进行了一些微生物学活性、免疫原性和毒力等方面的研究。
Recombinant vaccinia virus as live vaccine expressing glycoprotein D of HSV-1 168 strain was constructed.The gD gene under the control of 7.5k promoter from plasmid pJSB1175 was inserted into TK gene of vaccinia virus Tian Tan strain,a genetically stable strain of recombinant vaccinia virus vHSB1175D was constructed. The HSV-1 gD protein could be detected on the surface of cells infected with recombinant vaccinia virus vHSB1175D.The expressed antigen specifically reacted with the McAb against gD of HSV-1 and Western blot showed its molecular weights is 50kD.The gD gene was identified in vHSB1175D virus DNA by Southern blot hybridization.Some biological activities of the vHSB1175D were characterized and the antibodies against HSV-1 gD were produced in rabbits vaccinated by vHSB1175D.The specificity and high titers of the antibodies in rabbit sera were evaluated by ELISA and neutralization assays.The vHSB1175D was also tested in mice and protection against virus challenge was observed.
出处
《病毒学报》
CAS
CSCD
北大核心
1997年第3期193-201,共9页
Chinese Journal of Virology
关键词
痘苗病毒载体
疱疹病毒
病毒疫苗
糖蛋白D
研制
Vaccinia vector
HSV virus vaccine
Recombinant vaccinia virus
HSV glycoprotein D
