Extensive genetic diversity of bat-borne polyomaviruses reveals inter-family host-switching events

Zhizhou Tan; Gabriel Gonzalez; Jinliang Sheng; Jianmin Wu; Fuqiang Zhang; Lin Xu; Peisheng Zhang; Aiwei Zhu; Yonggang Qu; Changchun Tu; Michael J. Carr; Biao He

Genomic DNA from each bat tissue sample was extracted manually using the TIANamp Table. 595 Oligonucleotide primers for inverse PCRs for amplification of the 5 kb PyV genomes were 596 design based on the partial VP1 sequences. Multiple pairs of hemi-nested degenerate primers 597 were subsequently designed for VP1 clusters which showed close phylogenetic relationships (see 598 S6 Table) . All ~5 kb products were amplified from bat genomic DNA by inverse PCR employing 599 the PrimeSTAR MAX DNA polymerase (Takara), purified by gel extraction (Axygen), cloned 600 into pCR-Blunt II-TOPO vector (Invitrogen) and bidirectionally sequenced by primer walking to 601 Phred quality scores > 30 by 3730xl DNA Analyzer (Genewiz). After removal of vector . CC-BY 4.0 International license is made available under a The copyright holder for this preprint (which was not peer-reviewed) is the author/funder. It . https://doi.org/10.1101/627158 doi: bioRxiv preprint sequences, the full genomes of each PyV were assembled by Geneious software using the partial 603 VP1 and 5 kb products.