Hepatīta B vīrusa morfoģenēzes pētījumi, izmantojot alfavīrusu ekspresijas sistēmu
Date
2015
Authors
Petaja, Guna
Journal Title
Journal ISSN
Volume Title
Publisher
Latvijas Universitāte
Abstract
Darba mērķis bija pētīt hepatīta B vīrusa (HBV) kapsīda, subvirālo daļiņu un virionu izveides nosacījumus, izmantojot alfavīrusa ekspresijas sistēmu. Inficēto BHK-21 šūnu preparātu analīze tika veikta ar ELISA, imunoblotu un imunocitoķīmiju. Tika veiksmīgi izveidoti rekombinanti SFV1-HBV vīrusi – PG – pilna genoma HBV vīruss un HBC – vīruss, ko veido kapsīda proteīni, kā arī pārbaudīti laboratorijas kolekcijā esošais saīsināta kapsīda proteīna ΔC vīruss, un apvalka proteīna 1/L vīruss. Veicot BHK-21 šūnu infekciju ar 1/L vīrusu kombinācijā ar HBC, PG vai ΔC vīrusiem, bija jāņem vērā alfavīrusu konkurence. Tika novēroti dažāda brieduma kapsīdi ar atšķirīgu nukleīnskābju saturu ko raksturo atšķirīgs pārvietošanās ātrums agarozes gēlā. Visefektīvākā virionu veidošanās tika novērota tikai pēc PG vīrusa ko-infekcijas ar 1/L vīrusu.
The aim of this Master’s thesis was to investigate the formation of capsids, virions and subviral particles of Hepatitis B virus (HBV), using alphavirus expression system. Infected BHK-21 cell lysates were analyzed with ELISA, Western blot and immunocytochemistry. Recombinant SFV1-HBV viruses – PG – full-length HBV genome virus and HBC – virus, composed of capsid proteins, were obtained and a shortened core protein virus ΔC and evelope protein virus 1/L were tested. Alpavirus competition had to be taken in consideration when infecting cells with 1/L virus in combination with HBC, PG or ΔC virus. Different types of mature capsids were observed, characterized by different migration in agarose gel. The most efficient virion formation was observed after coinfection with PG and 1/L viruses.
The aim of this Master’s thesis was to investigate the formation of capsids, virions and subviral particles of Hepatitis B virus (HBV), using alphavirus expression system. Infected BHK-21 cell lysates were analyzed with ELISA, Western blot and immunocytochemistry. Recombinant SFV1-HBV viruses – PG – full-length HBV genome virus and HBC – virus, composed of capsid proteins, were obtained and a shortened core protein virus ΔC and evelope protein virus 1/L were tested. Alpavirus competition had to be taken in consideration when infecting cells with 1/L virus in combination with HBC, PG or ΔC virus. Different types of mature capsids were observed, characterized by different migration in agarose gel. The most efficient virion formation was observed after coinfection with PG and 1/L viruses.
Description
Keywords
Bioloģija , PG , HBC , ΔC , 1/L , imunoblots