An analysis of the effect of the internal ribosome entry site of the encephalomyocarditis virus on the expression of the second gene in the bicistronic matrix in neurons of primary hippocampal cultures
- Авторлар: Petrovskaya L.E.1,2, Shtefanyuk V.S.3, Balaban P.M.4,5, Ostrovsky M.A.5,6, Malyshev A.Y.4,7
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Мекемелер:
- Pirogov Russian National Research University
- Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
- Maastricht University
- Institute of Higher Nervous Activity and Neurophysiology
- Moscow State University
- Emanuel Institute of Biochemical Physics
- Aff7
- Шығарылым: Том 11, № 4 (2017)
- Беттер: 277-281
- Бөлім: Experimental Articles
- URL: https://ogarev-online.ru/1819-7124/article/view/211328
- DOI: https://doi.org/10.1134/S1819712417040067
- ID: 211328
Дәйексөз келтіру
Аннотация
Molecular biological experiments sometimes require expression of two or more genes in a single cell with an accurate ratio between their expression levels. One of the methods to provide this control is the use of the internal ribosome entry site (IRES) from the encephalomyocarditis virus as a separating insert between two target genes in the expression vector. Previously, it was shown that the efficacy of translation of the gene after IRES varies considerably in a range from 6 to 100% depending on the cell type. In neurons, the exact ratio between the expression levels of genes that are located before and after the IRES in the expression vector is unknown. Here, we analyzed the ratio between the amounts of products of the first and second genes located before and after the IRES in a plasmid that was used to transfect neurons in a primary hippocampal culture. We created two plasmid vectors that contain genes of the yellow (Venus) and red (mCherry) fluorescent proteins in different orders, which are separated by the IRES. We found that the unmodified IRES sequence of the encephalomyocarditis virus decreases the expression in the second cistron by a factor of 2.7 in a primary culture of hippocampal neurons. These data will help us to use currently available libraries of mutant IRES sequences for accurate control of the relationships between the expression of different target genes in neurons.
Негізгі сөздер
Авторлар туралы
L. Petrovskaya
Pirogov Russian National Research University; Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry
Email: malyshev@ihna.ru
Ресей, Moscow; Moscow
V. Shtefanyuk
Maastricht University
Email: malyshev@ihna.ru
Нидерланды, Maastricht
P. Balaban
Institute of Higher Nervous Activity and Neurophysiology; Moscow State University
Email: malyshev@ihna.ru
Ресей, Moscow; Moscow
M. Ostrovsky
Moscow State University; Emanuel Institute of Biochemical Physics
Email: malyshev@ihna.ru
Ресей, Moscow; Moscow
A. Malyshev
Institute of Higher Nervous Activity and Neurophysiology; Aff7
Хат алмасуға жауапты Автор.
Email: malyshev@ihna.ru
Ресей, Moscow; ul. Butlerova 5a, Moscow, 117865
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