Инфекция и иммунитет. 2014; 4: 229-234
СОЗДАНИЕ ГИБРИДНЫХ РЕКОМБИНАНТНЫХ БЕЛКОВ НА ОСНОВЕ БЕЛКОВ VP6 И VP8 РОТАВИРУСА ЧЕЛОВЕКА ГРУППЫ А
https://doi.org/10.15789/2220-7619-2014-3-229-234Аннотация
Ротавирусная инфекция является одной из наиболее распространенных причин детских энтеритов. Инфицирование этим вирусом зачастую приводит к выраженной дегидратации организма ребенка. Обезвоживание организма сопровождает многие инфекционные заболевания и является самой распространенной причиной смертности. На сегодняшний день самым эффективным способом профилактики ротавирусной инфекции считается своевременная вакцинация. Однако вакцинация проводится с применением живых аттенуированных вакцин, что может приводить к развитию различных осложнений. Создание кандидатной вакцины в этой работе осуществлялось на основе рекомбинантных гибридных белков, являющихся активными агентами при развитии протективного иммунитета против ротавируса. В ходе исследования созданы гибридные белки VP6VP8 и FliCVP6VP8, подобран оптимальный протокол индукции генов, кодирующих данные белки. Белки VP6VP8 и FliCVP6VP8 наработаны и очищены с использованием металлоаффинной хроматографии.
Список литературы
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4. Dennehy H.P. Rotavirus Vaccines: an overview. Clin. Microbiol. Rev., 2008, pp. 198–208.
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Russian Journal of Infection and Immunity. 2014; 4: 229-234
DEVELOPMENT OF FUSION RECOMBINANT PROTEINS BASED ON VP6 AND VP8 OF HUMAN ROTAVIRUS A
https://doi.org/10.15789/2220-7619-2014-3-229-234Abstract
Rotaviruses are the most frequent cause of children enteritis. This infection often leads to severe dehydration of organism. Many infectious diseases are accompanied by fluid loss being the most common cause of death. Nowadays an early vaccination is considered to be the most effective way for prevention of rotavirus infection. However attenuated alive vaccines are used for this purpose which can result in different complications. The candidate vaccine presented in this study was designed on the basis of recombinant fusion proteins — the crucial active agents for development of immune response against rotaviruses. As part of the study the following steps were taken: the design of fusion proteins VP6VP8 and FliCVP6VP8; protocol adjustment for induction of genes encoding these proteins expression, which yields in high level of the protein with minor expenditures; production and purification of recombinant proteins VP6VP8 and FliCVP6VP8 with metal affinity chromatography.
References
1. Balaram P., Kien P.K., Ismail A. Toll-like receptors and cytokines in immune responses to persistent mycobacterial a 1. nd Salmonella infections. Int. J. Med. Microbiol., 2009, vol. 299, no. 3, pp. 177–185.
2. Bishop R.F., Davidson G.P., Holmes I.H., RuckB.J. Virus particles in epithelial cells of duodenal mucosa from children with acute non-bacterial gastroenteritis. Lancet 2, 1973, pp. 1281–1283.
3. Bruijning-Verhagen P., Mangen J.M.-J., Felderhof M., Hartwig G.N., van Houten M., Winkel L., de Waal J.W., Bonten J.M.M. Targeted rotavirus vaccination of high-risk infants; a low cost and highly cost-effective alternative to universal vaccination. BMC Medicine, 2013, vol. 11, p. 112.
4. Dennehy H.P. Rotavirus Vaccines: an overview. Clin. Microbiol. Rev., 2008, pp. 198–208.
5. Flewett T.H., Bryden A.S., Davies H.A. Virus particles in gastroenteritis. Lancet, 1973, vol. 302, iss. 7844, p. 1497.
6. Invitrogen. Ni-NTA purification system. User manual. Catalog nos. K950-01, K951-01, K952-01, K953-01, K954-01, R901-01, R901-10, R 901-15. Version C. 25-0496, 2006, 32 p.
7. Laemmli U.K. Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4. Nature, 1970, vol. 227, pp. 680–685.
8. Majumder K. Ligation-free gene synthesis by PCR: synthesis and mutagenesis at multiple loci of a chimeric gene encoding OmpA signal peptide and hirudin. Gene, 1992, vol. 110, no. 1, pp. 89–94.
9. Patton J.T. Rotavirus diversity and evolution in the post-vaccine world. Discovery Med., 2012, vol. 13, no. 68, pp. 85–97.
10. Schnagl R.D., Holmes I.H. Characteristics of the genome of human infantile enteritis virus (Rotavirus). J. Virol., 1976, vol. 19, no. 1, pp. 267–270.
11. Studier F.W. Protein production by auto-induction in high density shaking cultures. Protein Expr. Purif., 2005, vol. 41, no. 1, pp. 207–234.
