Kaposi's sarcoma-associated herpesvirus (KSHV) is causally related to Kaposi's sarcoma (KS), primary effusion lymphoma (PEL) and a proportion of cases of multicentric Castleman's disease (MCD). The ORF73 p...Kaposi's sarcoma-associated herpesvirus (KSHV) is causally related to Kaposi's sarcoma (KS), primary effusion lymphoma (PEL) and a proportion of cases of multicentric Castleman's disease (MCD). The ORF73 protein was cloned into pQE80L-orf73 and expressed in E.coli and purified. The expressed recombinant ORF73 was identified by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). A protein of about 27 kDa was expressed as expected. Western Blotting showed that the purified recombinant ORF73 reacted with KSHV positive serum. The immunogenicity of the recombinant ORF73 was further analysed by ELISA and the optimal conditions were determined. The ORF73 ELISA was used to compare the KSHV seroprevalence between Hubei and Xinjiang Han people. The Han people in Xinjiang have significantly higher KSHV seroprevalence than their counterparts in Hubei (6.7% vs 2.9%, P = 0.005).展开更多
The ORFK8.1 of Kaposi＇s sarcoma associated-herpesvirus （KSHV） was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf KS.1 was induced by isopropyl-b-D-thiogala...The ORFK8.1 of Kaposi＇s sarcoma associated-herpesvirus （KSHV） was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf KS.1 was induced by isopropyl-b-D-thiogalactopyranoside （IPTG）. The fusion protein was purified by chromatyography. The expressed protein and its purified product were identified by sodium dodecyl sulfate-polyacrylamide gel eletrophoresis （SDS-PAGE）. SDS-PAGE showed that a protein of 26 kDa was visualized as expected. A western blot assay was established to analyze the immunogenicity of purified recombinant ORFKS. 1 protein. The optimal condition of the recombinant ORFKS. 1 ELISA assay was confirmed： the concentration of antigen was 5 μg/mL, the dilution of serum was 1：200. We used the ELISA method to investigate the recombinant ORF KS. 1 protein＇s specificity, the data showed that the specificity of ORF KS.1 to detect KSHV was 100%. At the same time, 560 sera samples from Hubei province were detected by using ORFKS. 1 ELISA to investigate KSHV seroprevalence in this region. The KSHV seroprevalence in Hubei province is shown to be 6.80%.展开更多
To purify the protein encoding the small capsid protein （SCP） of KSHV and analyze its immunogenicity, the carboxyl terminus of orf65 of Kaposi＇s sarcoma associated-herpesvirus （KSHV） was expressed in a prokaryoti...To purify the protein encoding the small capsid protein （SCP） of KSHV and analyze its immunogenicity, the carboxyl terminus of orf65 of Kaposi＇s sarcoma associated-herpesvirus （KSHV） was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf65 was induced by isopropyl-13-D-thiogalactopyranoside （IPTG） and the fusion protein was purified by chromatography. The expressed protein and its purified product were identified by sodium dodecyl sulfate- polyacrylamide gel electrophoresis （SDS-PAGE） and showed that 9 kDa was the expected size of the purified orf65 protein. The antiserum was produced in rabbit which was immunized by purified orf65 protein. An ELISA assay was established to analyze the immunogenicity of the purified orf65 protein. The ELISA analysis demonstrated that orf65 protein has strong immune activity, and the immune activity of polyclonal antibody against orf65 was more than 4 fold higher than that in the serum of the non-immunized rabbit. These results demonstrate that purified orf65 protein has very strong immunogenicity and can be used in screening KSHV infection in the general population using ELISA.展开更多
Kaposi's sarcoma-associated herpesvirus(KSHV)is the primary etiological agent of Kaposi's sarcoma,primary effusion lymphoma and muticentric Castleman's disease.In common with the other herpesviruses,KSHV e...Kaposi's sarcoma-associated herpesvirus(KSHV)is the primary etiological agent of Kaposi's sarcoma,primary effusion lymphoma and muticentric Castleman's disease.In common with the other herpesviruses,KSHV exhibits both latent and lytic life cycles,both of which are characterized by distinct gene expression profiles and programs.KSHV encodes proteins which play essential roles in the inhibition of host adaptive and innate immunity,the inhibition of apoptosis,and the regulation of the cell cycle.KSHV also encodes several proteins which have transforming and intrcellular signalling activity.展开更多
MicroRNAs (miRNAs) play important roles in eukaryotes,plants and some viruses. It is increasingly clear that miRNAs-encoded by viruses can affect the viral life cycle and host physiology. Viral miRNAs could repress th...MicroRNAs (miRNAs) play important roles in eukaryotes,plants and some viruses. It is increasingly clear that miRNAs-encoded by viruses can affect the viral life cycle and host physiology. Viral miRNAs could repress the innate and adaptive host immunity,modulate cellular signaling pathways,and regulate the expression of cellular and viral genes. These functions facilitate viral acute and persistent infections,and have profound effects on the host cell survival and disease progression. Here,we discuss the miRNAs encoded by herpesviruses,and their regulatory roles involved in virus-host interactions.展开更多
基金Supported by the Research grants from Mega Scientific Project for HIV in China (2008ZX-10001-002)National Natural Science Foundation of Hubei Province (2008CDA013)+1 种基金the Scientific Research Foundation for the Returned Overseas Chinese Scholars,State Education Ministry, and National Institutes of Health (DE017333)Open Research Fund Program of the State Key Laboratory of Virology of China (2010012)
文摘Kaposi's sarcoma-associated herpesvirus (KSHV) is causally related to Kaposi's sarcoma (KS), primary effusion lymphoma (PEL) and a proportion of cases of multicentric Castleman's disease (MCD). The ORF73 protein was cloned into pQE80L-orf73 and expressed in E.coli and purified. The expressed recombinant ORF73 was identified by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE). A protein of about 27 kDa was expressed as expected. Western Blotting showed that the purified recombinant ORF73 reacted with KSHV positive serum. The immunogenicity of the recombinant ORF73 was further analysed by ELISA and the optimal conditions were determined. The ORF73 ELISA was used to compare the KSHV seroprevalence between Hubei and Xinjiang Han people. The Han people in Xinjiang have significantly higher KSHV seroprevalence than their counterparts in Hubei (6.7% vs 2.9%, P = 0.005).
基金Supported by the Knowledge Innovation Program of the Chinese Academy of Sciences Chinese Academy of Sciences (0702121YJ1)Open Research Fund Program of the State Key Laboratory of Virology of China (2007013)
文摘The ORFK8.1 of Kaposi＇s sarcoma associated-herpesvirus （KSHV） was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf KS.1 was induced by isopropyl-b-D-thiogalactopyranoside （IPTG）. The fusion protein was purified by chromatyography. The expressed protein and its purified product were identified by sodium dodecyl sulfate-polyacrylamide gel eletrophoresis （SDS-PAGE）. SDS-PAGE showed that a protein of 26 kDa was visualized as expected. A western blot assay was established to analyze the immunogenicity of purified recombinant ORFKS. 1 protein. The optimal condition of the recombinant ORFKS. 1 ELISA assay was confirmed： the concentration of antigen was 5 μg/mL, the dilution of serum was 1：200. We used the ELISA method to investigate the recombinant ORF KS. 1 protein＇s specificity, the data showed that the specificity of ORF KS.1 to detect KSHV was 100%. At the same time, 560 sera samples from Hubei province were detected by using ORFKS. 1 ELISA to investigate KSHV seroprevalence in this region. The KSHV seroprevalence in Hubei province is shown to be 6.80%.
基金Knowledge Innovation Program of the Chinese Academy of Sciences(0702121Y-J1)
文摘To purify the protein encoding the small capsid protein （SCP） of KSHV and analyze its immunogenicity, the carboxyl terminus of orf65 of Kaposi＇s sarcoma associated-herpesvirus （KSHV） was expressed in a prokaryotic expression system. The expression of recombinant E.coli containing pQE-80L-orf65 was induced by isopropyl-13-D-thiogalactopyranoside （IPTG） and the fusion protein was purified by chromatography. The expressed protein and its purified product were identified by sodium dodecyl sulfate- polyacrylamide gel electrophoresis （SDS-PAGE） and showed that 9 kDa was the expected size of the purified orf65 protein. The antiserum was produced in rabbit which was immunized by purified orf65 protein. An ELISA assay was established to analyze the immunogenicity of the purified orf65 protein. The ELISA analysis demonstrated that orf65 protein has strong immune activity, and the immune activity of polyclonal antibody against orf65 was more than 4 fold higher than that in the serum of the non-immunized rabbit. These results demonstrate that purified orf65 protein has very strong immunogenicity and can be used in screening KSHV infection in the general population using ELISA.
文摘Kaposi's sarcoma-associated herpesvirus(KSHV)is the primary etiological agent of Kaposi's sarcoma,primary effusion lymphoma and muticentric Castleman's disease.In common with the other herpesviruses,KSHV exhibits both latent and lytic life cycles,both of which are characterized by distinct gene expression profiles and programs.KSHV encodes proteins which play essential roles in the inhibition of host adaptive and innate immunity,the inhibition of apoptosis,and the regulation of the cell cycle.KSHV also encodes several proteins which have transforming and intrcellular signalling activity.
基金The Knowledge Innovation Program of the Chinese Academy of Sciences Chinese Academy of Sciences (0702121YJ1)Open Research Fund Program of the State Key Laboratory of Virology of China(2007013)+1 种基金National Science Foundation of China (A Type B Outstanding Abroad Young Scientist Award)National Institutes of Health (CA096512,CA124332,CA119889 and DE017333)
文摘MicroRNAs (miRNAs) play important roles in eukaryotes,plants and some viruses. It is increasingly clear that miRNAs-encoded by viruses can affect the viral life cycle and host physiology. Viral miRNAs could repress the innate and adaptive host immunity,modulate cellular signaling pathways,and regulate the expression of cellular and viral genes. These functions facilitate viral acute and persistent infections,and have profound effects on the host cell survival and disease progression. Here,we discuss the miRNAs encoded by herpesviruses,and their regulatory roles involved in virus-host interactions.
