Contents
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2021 year, V. 7, №4
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Plant genetics

N.A. Vinichenko, E.A. Salina Gliadins: structure and toxic epitopes (in russian)
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Abstract
Abstract: This review shows features of the gliadins structure and the distribution of epitopes associated with celiac disease, among gliadins of different types. Several priority approaches were considered to accelerate the development of wheat varieties with an improved composition of gluten, safe for health.
Key words: wheat; gluten; gliadins structure; celiac disease epitopes.
For citation: Vinichenko N.A., Salina E.A. Gliadins: structure and toxic epitopes. Pisma v Vavilovskii Zhurnal Genetiki i Selektsii = Letters to Vavilov Journal of Genetics and Breeding. 2021;7(4):163-171. DOI 10.18699/LettersVJ2021-7-21 (in Russian)
Abstract
Abstract: Analysis of the expression of the Q gene (spelt factor), controlling the main agronomic traits by which a human had selected wheat (Triticum L.) during domestication and breeding, showed that the formation of the ear morphology of cultivated wheat species really depends on the level of expression of this gene. Based on the results of these investigation, it was found that in addition to a violation in the 329 Q position of the protein, its function and morphology of the wheat ear are also influenced by other factors and genes.
Key words: wheat; domestication; fragility.
For citation: Golovnina K.A., Blinov A.G., Mayorov V.I., Watanabe N., Goncharov N.P. Expression analysis of Q gene, controlling of the important domestication and breeding wheat traits. Pisma v Vavilovskii Zhurnal Genetiki i Selektsii = Letters to Vavilov Journal of Genetics and Breeding. 2021;7(4):172-175. DOI 10.18699/LettersVJ2021-7-22 (in Russian)

Biotechnology

Abstract
Abstract: Present day transgenesis tools make it possible to carry out quite complex modifications of animal genomes for different goals. In biotechnology, transgenic animals are generated to modify the composition of milk, including antibacterial properties, lipid composition, or flavor. One of the promising directions in this area is the overexpression of β-galactosidases, enzymes that hydrolyze lactose, to remove this disaccharide from the milk of farm animals. In our work, we investigate an approach with the expression of the transgalactasylating enzyme of the lactic acid bacterium Lactobacillus delbrueckii. In theory, the synthesis of this enzyme in the mouse mammary gland should lead to the disappearance of lactose from milk, due to its conversion into galactooligosaccharides. We created two transgenic animal lines to test this hypothesis. At the first stage, we tested variants of genetic constructs, including Golgi localization signals, and selected two variants of the β-galactosidase transgene under the constitutive and casein promoters to obtain transgenic mouse strains by pronuclear microinjection. For each of the constructs, four transgenic founders were obtained. We carried out a preliminary analysis of the characteristics of transgenic lines (transgene copy number, expression, inheritance of the transgene) and collected milk from lactating females for further research.
Key words: transgenesis; milk; lactose; galactooligosaccharides; Golgi apparatus.
For citation: Smirnov A.V., Korablev A.N., Yunusova A.M., Shnaider T.A., Serova I.A. Generating transgenic mice for investigating direct lactose conversion into galactooligosaccharides in mammary gland. Pisma v Vavilovskii Zhurnal Genetiki i Selektsii = Letters to Vavilov Journal of Genetics and Breeding. 2021;7(4):196-193. DOI 10.18699/LettersVJ2021-7-23 (in Russian)
Abstract
Abstract: In this study we cloned a cellulose from Penicillium sp. ‘occitanis’ and analyzed its properties. The gene was extracted from the NCBI database. We constructed a pPZL-4xBGLU-Pocc plasmid carrying four copies of a construction capable of expressing the P. sp. ‘occitanis’ cellulase. The plasmid was inserted into the Komagataella phaffii T07 yeast strain under the control of AOX1 promoter and terminator. A total of four copies of the expressed construction was cloned. As the result, we obtained the K. phaffii T07 BGLU-4xPоcc strain. The maximum expression level of the strain was ~4 000 000 U per 1 l of culture media in a 5 l bioreactor. The obtained protein was seen as a ~39.5 kDa band on the gel, which accords with its calculated mass. It also formed a smear from about 42 to 70 kDa, which probably indicates its glycosylation during the expression in K. phaffii. The temperature optimum of the enzyme was at 55–65 °C, and рН at ~1.5–2. The enzyme retains at least 50 % activity at pH ranging from 1.5 to 6, and at 30 to 80 °C. Below 75 °C, the protein remains active for 4 h. At 85 °С, it loses 50 % of its activity after 4 h. The enzyme is stable in pH ranging from 4 to 7.5.
Key words: cellulase; carbohydrase; endo-β-1,4-glucanase (EC 3.2.1.4); Penicillium sp. ‘occitanis’; Pichia pastoris; Komagataella phaffii; biotechnology; cultivation.
Acknowledgements: The study was funded by the Ministry of Science and Higher Education of the Russian Federation as a part of the comprehensive project on high-tech industry “Creation of high-tech production of high-quality plant food proteins” (The agreement on the provision of subsidies from the federal budget for the development of cooperation of a state scientific institution and organization the real sector of the economy in order to implement a comprehensive project for the creation of high-tech industry No. 075-11-2020- 036 from 15.12.2020) in the framework of the Decree of the Government of the Russian Federation of April 9, 2010 No. 218 on the basis of the ICG SB RAS. We thank ICG SB RAS Core Facility of Proteomic and metabolomic profiling of microorganisms (supported by project No. 0259-2021-0010).
For citation: Rozanov A.S., Voskoboev M.E., Bogacheva N.V., Korzhuk A.V., Shlyahtun V.N., Mescheryakova I.A., Romancev V.A., Bochkov D.V., Zadorozhnyy A.V., Peltek S.E. Cloning and expression of a cellulase gene from Penicillium sp. ‘occitanis’ in Komagataella phaffii T07. Pisma v Vavilovskii Zhurnal Genetiki i Selektsii = Letters to Vavilov Journal of Genetics and Breeding. 2021;7(4):194-200. DOI 10.18699/LettersVJ2021-7-24 (in Russian)

Letters to the Editor

Abstract
For citation: Glazko V.I. The scientific component of breeding: questions raised in the special issue of the Vavilov Journal of Genetics and Breeding, 2021;25(4). Pisma v Vavilovskii Zhurnal Genetiki i Selektsii = Letters to Vavilov Journal of Genetics and Breeding. 2021;7(4):201-205. DOI 10.18699/LettersVJ2021-7-25 (in Russian)