Expression of the Nucleotide Sequence for the M2e Peptide of Avian Influenza Virus in Tobacco Transgenic Plants

Автор: I.V. Tarasenko, A.I. Taranov, A.P. Firsov, and S.V. Dolgov

Страница: 18-25

Expression of the Nucleotide Sequence for the M2e Peptide of Avian Influenza Virus in Tobacco Transgenic Plants

Biotekhnologiya, 2012, N 4, P. 18-25

UDC 575.113.1:577.218:633.71]:615.371

Section:  “Producers, Biology, Selection, and Gene Engineering”

I.V. Tarasenko ^{1, 2, *},  A.I. Taranov ²,  A.P. Firsov ^{1, 2, *},  and  S.V. Dolgov ^{1, 2}

¹  The All-Russian research Institute for Agricultural Biotechnology,  127550, Moscow Russia

²  The Shemyakin-and-Ovchinnikov Insitute for Bioorganic Chemistry, Russ. Acad. Sci., Pushchino branch,  142290, Pushchino, Moskovskaya oblast Russia

e-mail:  aleksey_firsov@mail.ru,  starassenko@rambler.ru

An M2e peptide of the H5N1 A/Chicken/Kurgan/05/2005 avian influenza virus has successfully been synthe­sized in transgenic tobacco plants. The N-terminal segment of M2 protein, 22, 30 or 43 amino acids in length including the M2e peptide (М122, М130 and М143 variants, respectively), was translationally fused with the N-terminus of β-glucuronidase in the plant expression vector of pBI121. The nucleotide sequence of the target fragment was synthesized by ligation from synthetic oligonucleotides, its codon composition being previously adapted for the expression in plants. Tobacco plants were successfully transformed with the obtained vectors (pBIM122, -130 и -143, respectivily). In the plants transformed with pBIM143, the fused protein М143-β-glucuronidase was not produced which is possibly a result of the occurrence of an M2 protein transmembrane domain (25—43 aa of M2) in this construct. In the pBIM122- and pBIM130-transformed plants, the target peptide M2e was expressed as a part of the fused proteins of М122-β-glucuronidase and М130-β-glucuronidase, respectively, as was detected by Western-blot analysis. These proteins occurred as bands of the expected size without apparent degradation. As a result, the M2e peptide of the H5N1 avian influenza virus was successfully synthesized for the first time in nuclear-transformed transgenic plants. The obtained results will be used in designing of transgenic plants-based edible anti-influenza vaccine.

Key words:  avian influenza,  edible vaccines,  molecular farming,  M2e peptide.

The full English version of the article was published in “Applied Biochemistry and Microbiology”, 2013, Vol. 49, Issue 8, pp. 695-701 as I. V. Tarasenko, A. I. Taranov, A. P. Firsov, S. V. Dolgov “Expression of the nucleotide sequence for the M2e peptide of avian influenza virus in transgenic tobacco plants”.

It is contained at the SpringerLink website:  http://link.springer.com/article/10.1134/S0003683813080061  and  at the Russian Scientific Electron Library website:  http://elibrary.ru/item.asp?id=21889067

DOI: 10.1134/S0003683813080061

08.05.2015, 1613 просмотров.