Hepatīta B core mutantu konstruēšana, ekspresija un attīrīšana strukturāliem pētījumiem un jaunu vakcīnu prototipu izstrādei

6533b82efe1ef96bd129412b

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Hepatīta B core mutantu konstruēšana, ekspresija un attīrīšana strukturāliem pētījumiem un jaunu vakcīnu prototipu izstrādei

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Darba mērķis bija konstruēt un attīrīt dažādus hepatīta B vīrusa (HBV) nukleokapsīda proteīna (HBc) mutantus (a) strukturāliem pētījumiem un (b) jaunu vakcīnu prototipu izstrādei. HBc dimēru mijiedarbības pētījumiem ar šūnas transportproteīniem un/vai kodola poru kompleksa proteīniem uz 145 aminoskābju garā HBc (HBcΔ145) bāzes tika konstruēti mutanti: N33→R, A36→Q un R127→E. Divi pēdējie tika veiksmīgi ekspresēti un attīrīti. Abi veidoja dimērus, bet ne nukleokapsīdas, tomēr A36→Q proteīns producējās labāk un tika iegūts ar augstāku tīrības pakāpi. Jaunu vakcīnu prototipu izstrādei tika pārbaudīta HBc mutanta E77→K spēja ķīmiski saistīt Laima boreliozes izraisītājas Borrelia burgdorferi ārējās virsmas proteīnu BB0689. Tika konstruēts E77→K mutants uz 176 aminoskābju gara HBc bāzes. Izveidotais konstrukts tika ekspresēts E. coli un nukleokapsīdu formā attīrīts no šūnām. Šī struktūra bija piemērota BB0689 proteīna ķīmiskai piesaistei, un ar himērajām daļiņām tika imunizētas peles.

year	journal	country	edition	language
2014-01-01

https://dspace.lu.lv/dspace/handle/7/18954

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