%0 Journal Article
%T Construction and immunogenicity of recombinant porcine parvovirus-like particles with somatostatin<br>展示生长抑素的猪细小病毒样颗粒构建及其免疫原性
%A Xuehua Zhang
%A Qisheng Zheng
%A Jin Chen
%A Gang Xue
%A Hongyan Hou
%A Jibo Hou
%A <br>张雪花
%A 郑其升
%A 陈瑾
%A 薛刚
%A 侯红岩
%A 侯继波
%J 生物工程学报
%D 2010
%I 
%X In order to obtain a virus-like particle vaccine both for porcine parvovirus (PPV) prevention and growth-promotion, VP2 gene of PPV NJ-a strain was amplified with PCR, and four copies of synthetic somatostatin gene were fused to the N-terminal of VP2 gene. The fused gene was cloned into pFast-HT A to construct the recombinant plasmid pFast-SS4-VP2, then the pFast-SS4-VP2 was transformed into DH10Bac competent cells and recombined with shuttle vector Bacmid, followed by identification with blue-white screening and PCR analysis for three cycles, and the positive recombinant was named as rBacmid-SS4-VP2. The positive Sf-9 cells were transfected with rBacmid-SS4-VP2 by Lipofectamine to produce recombinant baculovirus. When the cytopathic effect (CPE) was obvious, the transfected Sf-9 cell was harvested, and the positive recombinant virus was named as rBac-SS4-VP2. The insertion for the target gene into baculovirus genome was confirmed with PCR. SDS-PAGE and Western blotting revealed that the calculated protein of approximately 68 kDa was in the expressed in the insect cells. The Sf-9 cells infected with rBac-SS4-VP2 were stained positive against PPV antibody using the indirect immunofluorescence assay (IFA). Moreover, the virus particle self-assembly was observed under electron microscopy. 90 four-week-old mice were immunized by the recombinant protein coupled with different adjuvants alhydrogel, IMS and oil. VP2-specific ELISA antibodies, PPV-specific neutralizing antibody, somatostatin antibody and growth hormone levels were examined to evaluate the immunogenicity of this virus like particle. Results indicated that mice groups immunized rSS4-VP2 protein with alhydrogel and IMS developed similar humoral immune response comparing with inactived PPV vaccine. Mice group immunized with rSS4-VP2 generated higher level of SS antibody and growth hormone comparing with negative control, mice receiving rSS4-VP2 with alhydrogel developed the highest antibody titre than all other groups, while the oil group developed the lowest antibody level. This study provides not only a new rout for production of safe and effective virus like particle subunit vaccine, but also the foundations for peptide presentation and multivalent subunit vaccine design.
%K porcine parvovirus
%K VP2
%K somatostatin
%K baculovirus
%K virus-like particles<br>猪细小病毒，VP2蛋白，生长抑素，杆状病毒，病毒样颗粒
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=86F37710D2D8EF0706DDF86AEE540AE3&yid=140ECF96957D60B2&vid=96C778EE049EE47D&iid=5D311CA918CA9A03&sid=C6E83222BCBFFFF8&eid=0AFD076159674A31&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=0