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OALib Journal期刊
ISSN: 2333-9721
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Expression of Recombinant Human Cytomegalovirus Fusion Proteins pp150/MDBP Fragments and its Application
人巨细胞病毒pp150蛋白片段与MDBP蛋白片段的融合表达及初步应用

Keywords: Human cytomegalovirus,fusion protein,prokaryotic expression,Western blot,protein chip
人巨细胞病毒
,融合蛋白,原核表达,Western,blot,蛋白芯片

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Abstract:

Human cytomegalovirus(HCMV) infection is an ubiquitous herpesvirus disease in human populations. It is rarely pathogenic to healthy adults,yet it may cause severe outcome to organ transplant recipients,the immunocompromised individuals and pregnant women. Using DNA from HCMV AD169 strain as template,the UL32 gene encoding pp150 protein fragment and the UL57 gene encoding MDBP protein fragment were amplified by PCR technique. After the construction of cloning vector pMD18-T-UL32,pMD18-T-UL57,pMD18-T-UL32/UL57 and expression vector pET-11a-UL32/UL57,the recombinant fusion proteins pp150/MDBP were induced with IPTG in BL21 host strain. The results showed that the relative molecular weight of recombinant fusion proteins pp150/MDBP is about 27kD,the product of fusion proteins takes 17.45% in the total proteins in host bacteria,the analytical result was positive to the fusion proteins pp150/MDBP via Western blot technique,while the purified recombinant fusion proteins have strong antigenicity detected by ELISA and protein chip compared with whole virus antigens from HCMV. It was demonstrated that when used for the detection of serum from the clinical patients it has the same detection rate compared with the whole virus antigen. It needs further research for application.

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