CpG oligonukleotīdu pakošana RGD tripeptīdu saturošajās vīrusveidīgajās daļiņās
Date
2015
Authors
Andersone, Santa
Journal Title
Journal ISSN
Volume Title
Publisher
Latvijas Universitāte
Abstract
Pasaulē ar hepatīta C vīrusu inficēto cilvēku skaits ir apmēram 170 miljoni, un pret HCV nav vakcīnas. Tāpēc ir svarīgi veikt pētījumus, kas saistīti ar vakcīnu izstrādi pret hepatīta C vīrusu.
Darba mērķis bija veikt CpG oligodezoksinukleotīdu pakošanu vīrusveidīgajās daļiņās, kur kā nesējs tika izmantots 176 as garš hepatīta B kora modificēts proteīns. Nesēja galvenais imunodominantais rajons saturēja RGD tripeptīdu, un tā C gals bija sapludināts ar HCV NS3 222 – 251 as garu fragmentu.
Tika attīrīti vairāki himēro HBc/HCV NS3 222 – 251 varianti. Tika veikta CpG oligodezoksinukleotīdu pakošana trīs dažādās ekspresijas plazmīdās. Pēc apstrādes ar DNāzi I tika konstatēts, ka vīrusveidīgo daļiņu saistītie CpG oligodezoksinukleotīdi atrodas daļiņu iekšpusē un ir pasargāti no DNāzes I. Visas ekspresijas plazmīdas uzrādīja līdzīgus rezultātus CpG oligodezoksinukleotīda iepakošanā.
The number of people infected by hepatitis C virus in the world is about 170 million, and there is no suitable vaccine against the HCV. Therefore it is important to carry out research that are connected to finding a vaccine for Hepatitis C virus. The aim of the work was to carry out a CpG oligodeoxynucleotide packaging into virus like particles, where as a carrier was used 176 aa long hepatitis B core modified protein with RGD tripeptide in the major immunodominant region and its C-terminal was fused with HCV NS 3 222 – 251 aa long fragment. Many chimer HBc176/HCV NS3 222 - 251 variants were purified. CpG oligodeoxynucleotide packing was made in three different expression plasmids. After the treatment with DNase I was found that virus like particle containing CpG oligodeoxynucleotides are located inside the particles and are protected from DNase I. All expression plasmids shown similar results in CpG oligodeoxynucleotides packaging.
The number of people infected by hepatitis C virus in the world is about 170 million, and there is no suitable vaccine against the HCV. Therefore it is important to carry out research that are connected to finding a vaccine for Hepatitis C virus. The aim of the work was to carry out a CpG oligodeoxynucleotide packaging into virus like particles, where as a carrier was used 176 aa long hepatitis B core modified protein with RGD tripeptide in the major immunodominant region and its C-terminal was fused with HCV NS 3 222 – 251 aa long fragment. Many chimer HBc176/HCV NS3 222 - 251 variants were purified. CpG oligodeoxynucleotide packing was made in three different expression plasmids. After the treatment with DNase I was found that virus like particle containing CpG oligodeoxynucleotides are located inside the particles and are protected from DNase I. All expression plasmids shown similar results in CpG oligodeoxynucleotides packaging.
Description
Keywords
Bioloģija , HCV , HBc , RGD , VLP , CpG