Whole Sequence Analysis And Expression Of The Protein Of Bovine Papillomavirus 1 Genotype SY-12 Strain In Southern Xinjiang

Bovine papillomatosis is a neoplastic disease caused by Bovine papillomavirus(BPV).The cattle was infected with the disease,mainly manifested by the prominent supernumerary tumor outside the body surface,and it was easily to cause cross infection through the contact between herds.Bovine papillomavirus type I and type II can be transmitted across different species through herd feeding,mainly due to infection of bovine papillomavirus in equine animals.Because of its host specificity,tissue limitation and difficulty in cell proliferation,the papillomavirus is currently at the molecular level.In this study,based on a sample genotyping identification of a suspected case of infected bovine papilloma in southern Xinjiang,the whole genome sequence of the bovine papillomavirus type 1 SY-12 strain and its structural characteristics were analyzed.The prokaryotic expression system of the SY-12 L1 gene and the E6 eukaryotic expression system were constructed to further study the genomes of bovine papilloma.It provides a theoretical basis for further studies on genotyping,traceability of epidemic sources,molecular genetic evolution and vaccine research of bovine papilloma.1.On the basis of clinical diagnosis and histopathological examination,the L1 gene fragment was amplified and sequenced successfully with the common primer MY11/MY09 of papillomavirus,and the nucleotide homology was compared and analyzed to determine that the infected animal was BPV-1 type bovine papillomavirus infection.2.The whole genome sequence of bovine papillomavirus type 1 SY-12 strain was sequenced by specific amplification primers and sequence primers.The full-length of the SY-12 strain was 7946 bp,with the functional gene structure of BPV,including the early coding region gene E6,E7,E1,E2,E4,E5 and late coding region genes L1 and L2,whichbelongs to the genus Delta genus bovine papillomavirus.3.The target fragment of 1488 bp was amplified with the specific primers of the L1 gene and the prokaryotic expression system pET32a-L1 of the BPV-1 L1 gene was constructed.The target fusion protein with a molecular weight of 75 KD was obtained after the optimization of the induction condition,which was in accordance with the expected protein size.4.The BPV-1 E6 gene was cloned successfully with the specific primers of E6 gene.The target gene was 414 BP.The E6 eukaryotic expression system pEGFP-N1-E6 was constructed and transfected into BHK-21 cells.The clear green fluorescence was visible under the inverted fluorescence microscope.