[Expression of recombinant human cytomegalovirus fusion proteins pp150/MDBP fragments and its application].
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 27 kD, 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.
Guo
Da-Dong
DD
Shandong Medicinal Biotechnology Center, Key Laboratory for Biotech-drugs of the Ministry of Health, China.
Gao
Xue-Qin
XQ
Han
Jin-Xiang
JX
Liu
Yi
Y
Zhang
Hua-Ning
HN
D003587:Cytomegalovirus; D004268:DNA-Binding Proteins / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D010446:Peptide Fragments / Q000097:blood / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D010750:Phosphoproteins / Q000737:chemistry*; D010957:Plasmids / Q000235:genetics; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000097:blood / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification; D014157:Transcription Factors / Q000737:chemistry*; D014763:Viral Matrix Proteins / Q000737:chemistry*
D004268:DNA-Binding Proteins; C066141:MBD1 protein, human; D010446:Peptide Fragments; D010750:Phosphoproteins; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; D014763:Viral Matrix Proteins; C056367:pp150 protein, Cytomegalovirus