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Recombinant human zona pellucida proteins ZP1, ZP2 and ZP3 co-expressed in a human cell line

Asian Journal of Andrology 2004年第1期6卷 3-13页

作者： MirjanaMartic erick.moses TimE.Adams DeYiLiu DebraA.Gook ClaireGarrett MarjorieE.Dunlop GordonH.W.Baker DepartmentofObstetricsandGynaecology，UniversityofMelbourne DepartmentofMedicine UniversityofMelbourne PregnancyResearchCenterRoyalWomen''sHospital ReproductiveServices RoyalWomen''sHospitalandMelbourneIVE CSIRODivisionofHealthScienceandNutrition MelbourneAustralia

Aim: To produce biologically active recombinant human (rh) ZP proteins in a human cell for use in sperm function tests. Methods: The human embryonic kidney cell line 293T was employed to produce rhZP1, rhZP2 and rhZP3 proteins individually and together by co-expression. Presence of these proteins in the culture medium and cell lysate was assessed by Western blotting analysis. The effect of the recombinant proteins on the human AR was assessed. Results: RhZP2 and rhZP3 were secreted into the culture medium, whereas rhZPl was found only in the cell lysate. Interestingly, when all zona pellucida proteins were co-expressed in the same cells, rhZPl was also secreted into the culture medium. However, despite the presence of all three ZP proteins in sufficient concentration and evidence of heavy glycosylation on gel electrophoresis, biological activity to induce the AR was not observed. Conclusion: RhZP1, rhZP2 and rhZP3 were successfully expressed in the human embryonic kidney cell line 293T. It appears that an interaction amongst these proteins may be required for release of rhZPl from the cell. Although this approach is not satisfactory for producing active human ZP proteins, it makes a significant contribution to the understanding of the structural and functional characteristics of the ZP proteins.

关键词： acrosome reaction glycosylation human cell line recombinant proteins zona pellucida

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Genetic Evidence of Middle East Respiratory Syndrome Coronavirus(MERS-Cov) and Widespread Seroprevalence among Camels in Kenya

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Virologica Sinica 2018年第6期33卷 484-492页

作者： Sheila Ommeh Wei Zhang Ali Zohaib Jing Chen Huajun Zhang Ben Hu Xing-Yi Ge Xing-Lou Yang moses Masika Vincent Obanda Yun Luo Shan Li Cecilia Waruhiu Bei Li Yan Zhu Desterio Ouma Vincent Odendo Lin-Fa Wang Danielle E.Anderson Jacqueline Lichoti erick Mungube Francis Gakuya Peng Zhou Kisa-Juma Ngeiywa Bing Yan Bernard Agwanda Zheng-Li Shi Institute for Biotechnology Research Jomo Kenyatta University of Agriculture and TechnologyNairobi 62000-00200Kenya CAS Key Laboratory for Special Pathogens and Biosafety Wuhan Institute of VirologyChinese Academy of SciencesWuhan 430071China College of Biology Hunan University410006China Department of Medical Microbiology University of NairobiNairobi 30197-00100Kenya Veterinary Services Department Kenya Wildlife ServiceNairobi 40241-00100Kenya Veterinary Research Institute Kenya Agriculture and Livestock Research OrganizationNairobi 57811-00200Kenya Veterinary Services Programme in Emerging Infectious Diseases Duke-NUS Medical SchoolSingapore 169857Singapore Directorate of Veterinary Services State Department of LivestockMinistry of AgricultureLivestock Fisheries and IrrigationNairobi 34188-00100Kenya Kenya Camel Association Nairobi 30095-00100Kenya Department of Zoology National Museums of KenyaNairobi 40658-00100Kenya

We describe the first genome isolation of Middle East respiratory syndrome coronavirus(MERS-CoV) in Kenya. This fatal zoonotic pathogen was first described in the Kingdom of Saudi Arabia in 2012. Epidemiological and molecular evidence revealed zoonotic transmission from camels to humans and between humans. Currently, MERS-CoV is classified by the WHO as having high pandemic potential requiring greater surveillance. Previous studies of MERS-CoV in Kenya mainly focused on site-specific and archived camel and human serum samples for antibodies. We conducted active nationwide cross-sectional surveillance of camels and humans in Kenya, targeting both nasal swabs and plasma samples from 1,163 camels and 486 humans collected from January 2016 to June 2018. A total of 792 camel plasma samples were positive by ELISA. Seroprevalence increased with age, and the highest prevalence was observed in adult camels(82.37%, 95%confidence interval(CI) 79.50–84.91). More female camels were significantly seropositive(74.28%, 95% CI 71.14–77.19)than male camels(P \ 0.001)(53.74%, 95% CI 48.48–58.90). Only 11 camel nasal swabs were positive for MERS-CoV by reverse transcription-quantitative PCR. Phylogenetic analysis of whole genome sequences showed that Kenyan MERSCoV clustered within sub-clade C2, which is associated with the African clade, but did not contain signature deletions of orf4 b in African viruses. None of the human plasma screened contained neutralizing antibodies against MERS-CoV. This study confirms the geographically widespread occurrence of MERS-CoV in Kenyan camels. Further one-health surveillance approaches in camels, wildlife, and human populations are needed.

关键词： Middle East respiratory syndrome coronavirus(MERS-CoV) One-health Public health Zoonosis Kenya

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