Viable Viruses in Neolithic Remains
Search for Viable Viruses and their Genetic Elements in Neolithic Remains of Gorny Altai and Old Cemeteries in the Permafrost Region of Russia
Tech Area / Field
- BIO-CGM/Cytology, Genetics and Molecular Biology/Biotechnology
3 Approved without Funding
State Research Center of Virology and Biotechnology VECTOR, Russia, Novosibirsk reg., Koltsovo
- Siberian Branch of RAS / Institute of Cytology and Genetics, Russia, Novosibirsk reg., Novosibirsk\nInstitute of Archaeology and Ethnography, Russia, Novosibirsk reg., Novosibirsk
- Universität München / Department of General Biology, Germany, Munich\nRockefeller University, USA, New York\nWHO Collaborating Center for Studies on the Ecology of Influenza in Animals, USA, TN, Memphis\nCenters for Disease Control and Prevention (CDC) / National Center for Infectious Diseases, USA, GA, Atlanta\nCentre for Applied Microbiology and Research, UK, Salisburg
Project summaryThe goal of the present project is the revealing of viral genomes and/or their fragments in the unique material - the Neolithic remains of the ancient man which were discovered in the region of Gorny Altai, as well as in the samples taken from corpses of men who had died in the epidemics of viral genesis at the end of the last century and were buried in the permafrost region of Russia. Basing on the studies of the primary sequences of viral genomes and/or their fragments and on their comparison with the presently available sequences of viral genomes, the evolutionary process of the viral genome will be reconstructed and the predictive model of the emerging of new viruses will be developed. Besides, the epidemic potential and danger of the cemeteries located in the permafrost region where those died of smallpox were buried will be comprehensively estimated and substantiated recommendations on the strategy for controlling the orthopoxviral infections distribution will be presented.
This objective will be solved basing on the method of degenerate oligonucleotide markers, which will be theoretically calculated in such a way as to reveal only fragments of viral genomes (precisely distinguishing them from the fragments of bacterial and eucaryotic origin). Such oligonucleotide primers will provide the possibility to perform the search for viral genomes and/or their fragments starting from the viral superfamilies; gradual decrease of the primers' degeneracy extent will provide the revealing of viral families, etc.
The following stages will be carried out during the fulfillment of the present Project:
1. Theoretical analysis of the peculiarities of viral genomes for selection of PCR primers structure.
2. Simultaneously the archive search of the descriptions of various epidemics of infectious diseases in the permafrost zone, Gorny Altai, and other Siberian regions will be carried out in attempt to choose the optimal place for expeditions aiming to take the samples from the corpses.
3. Expedition to the chosen region of Siberia for sampling the bioremnants.
4. Isolation of nucleic acids preparations from paleoremnants. PCR amplification of the human genome fragments and subsequent amplification of the viral genome fragments. Creation of the bank of DNA samples (virus carriers, mixtures of the cloned fragments, etc.) for development of a control system while molecular genetic analysis of bioremnants. Sampling of the modern native population in the region of bioremnant revelation.
5. Study of the viral virus viability after isolation from the corpses in the permafrost region and its complex epidemiological evaluation.
6. Molecular genetic analysis of the nucleic acids isolated from the bioremnants for the presence of viral genomes comprising (I) amplification of viral genomes either through PCR, or through the direct cloning of isolated DNA; (ii) identification of presumable viral genome fragments by various molecular genetic approaches.
7. Computer analysis of the molecular genetic data obtained for constant control of specific implement of the amplified and cloned material.
8. Molecular genetic analysis of anthropologically informative genetic markers for biosamples and DNA preparations of modern native population to determine the possible evolutionary history of the inpiduals under study. Polymorphism analysis of mitochondria DNA and several informative nuclear hypervariable markers.
9. Reconstruction of the evolutionary process of the viral genome. Comprehensive comparison of the identified viral genome fragments obtained with the available sequences of viral genomes. On the basis of the data obtained, the conclusions will be made concerning the regularities of viral genome evolution and prognostic models of novel viruses emergence will be developed; data on viability of the revealed viruses will be obtained.
Results of the present Project will be not only of basic importance, but will also lay the basis for enhancing the efficacy of the prognostic and prophylactic measures.
Participation of the foreign collaborators is desirable for consultations and discussion while obtaining of the initial preparations for subsequent molecular studies and while carrying out the theoretical analysis of the results obtained at the final stage of the project fulfillment while reconstructing the evolutionary process of viruses.