摘要
为研究牛细小病毒(BPV)VP2蛋白免疫学相关功能,本研究采用PCR扩增得到BPV-1VR767株VP2基因片段,测序后利用生物学软件分析VP2蛋白的抗原表位,选取抗原表位富集的3个基因片段VP2-1(100 bp～789 bp),VP2-2(898 bp～1 353 bp)和VP2-3(1 450 bp～1 899 bp)分别克隆至原核表达载体pET-28a中,并转化E.coli中进行诱导表达。表达的重组蛋白经western blot和间接ELISA鉴定;用纯化的重组蛋白分别免疫小鼠,并用ELISA检测小鼠多克隆抗体特异性。结果显示:表达的3种重组蛋白分子量分别约为30.2 ku、22.9 ku和22.7 ku,均可以被BPV阳性血清识别。间接ELISA结果表明,3种重组蛋白均能够有效诱导抗体反应。其中,重组VP2-3诱导产生的抗体效价最高,与阳性血清的结合能力最强。该研究结果为研究VP2蛋白功能、建立BPV检测方法提供了基础材料。
To identify the antigenicity in different VP2 regions of bovine parvovirus-1 （BPV-1）, the VP2 gene of BPV-1 VR767 strain was amplified by PCR and sequenced. Three encoding immunodominant regions of BPV VP2-1 （aa34 - aa263）, VP2-2 （aa300 - aa451） and VP2-3 （aa484 - aa633） were analyzed by bioinformatics softwares and subcloned into pET28a for expressions in E.coli, which were 30.2 ku, 22.9 ku and 22.7 ku, respectively, identified by Western blot. Furthermore, the mice were immunized with the truncated recombinant proteins to prepare the antisera, respectively. Western blot and indirect ELISA analysis showed that three truncated proteins specifically reacted with antiserum against BPV. In addition, all of the three proteins were able to induce antibody responses in mice and VP2-3 induced the highest antibody titers. The expressions of recombinant protein and truncated proteins of VP2 provide the basic for further study the functions of VP2 protein and establish BPV detection methods.
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2013年第7期546-549,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
国家863课题(2011AA10A213)
校博士启动基金(2009RC03)
关键词
牛细小病毒
VP2基因
表达
抗原性分析
bovine parvovirus
VP2 gene
expression
analysis of antigenicity
