17 research outputs found
The Plant Nutrition from the Gas Medium in Greenhouses: Multilevel Simulation and Experimental Investigation
ΠΡΠΎΠ±Π΅Π½Π½ΠΎΡΡΠΈ ΠΏΡΠΎΠ±ΠΎΠΏΠΎΠ΄Π³ΠΎΡΠΎΠ²ΠΊΠΈ Π»ΠΈΠ·Π°ΡΠΎΠ² Π΄Π»Ρ ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΡ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ Π²ΡΠ΄Π΅Π»Π΅Π½ΠΈΡ Π±Π΅Π»ΠΊΠΎΠ²ΡΡ ΠΏΠ°ΡΡΠ½Π΅ΡΠΎΠ² ΡΠ΅Π»Π΅Π²ΡΡ Π±Π΅Π»ΠΊΠΎΠ², ΠΊΠΎΠ΄ΠΈΡΡΠ΅ΠΌΡΡ Π³Π΅Π½Π°ΠΌΠΈ 18-ΠΎΠΉ Ρ ΡΠΎΠΌΠΎΡΠΎΠΌΡ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ°
The aim of this work was to test modifications of the standard protocol for the sample preparation of cell/tissue lysate before performing the affinity isolation of lysate protein partners for the target protein (bait protein) which is covalently immobilized on an inert sorbent (e.g. BrCN-, SH-Sepharose 4B) or a carrier (e.g. paramagnetic nanoparticles). The series of our previous works on applying the approach to direct molecular fishing procedure with combination of affinity chromatography and LC-MS/MS analysis using a number of proteins, encoded by the genes of human chromosome 18, have shown that there are at least two problems affecting the specificity and the effectiveness of this procedure. These include: (i) redundancy of the background proteins in the eluates from an affinity sorbent (carrier) due to isolation of multiprotein complexes βlabeledβ with a direct protein partner which binds with a bait protein immobilized on the sorbent; (ii) low enrichment of the eluates with appropriate protein partners due to the fact that some direct protein partners in the lysate exist in stable βwild typeβ complexes with the bait protein itself. This means that latter group of protein partners will not be sufficiently isolated from lysate. Therefore, in order to increase the specificity and efficiency of affinity isolation of protein partners for the bait protein, we modified the standard protocol of lysate preparation and the preliminary step on dissociation of lysate protein complexes was added. Several model experiments for the choice of regeneration solution, assessment of their efficiency in the dissociation of lysate protein complexes as well as the stability and binding capacity of proteins were performed under the control of surface plasmon resonance (SPR) biosensor Biacore 3000 using HepG2 cell lysate. It was shown that acid treatment and incubation of the cell lysate for one min on ice (final lysate dilution 20 times) and subsequent neutralization (pH shift from 2.0 to 7.4) resulted in maximal dissociation of the lysate protein complexes without significant negative effects on the protein-protein interactions tested.Π¦Π΅Π»ΡΡ ΡΠ°Π±ΠΎΡΡ Π±ΡΠ»ΠΎ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°Π»ΡΠ½ΠΎΠ΅ ΡΠ΅ΡΡΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ ΠΌΠΎΠ΄ΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ ΡΡΠ°Π½Π΄Π°ΡΡΠ½ΠΎΠ³ΠΎ ΠΏΡΠΎΡΠΎΠΊΠΎΠ»Π° ΠΏΡΠΎΠ±ΠΎΠΏΠΎΠ΄Π³ΠΎΡΠΎΠ²ΠΊΠΈ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠ³ΠΎ/ ΡΠΊΠ°Π½Π΅Π²ΠΎΠ³ΠΎ Π»ΠΈΠ·Π°ΡΠ° ΠΏΠ΅ΡΠ΅Π΄ Π²ΡΠΏΠΎΠ»Π½Π΅Π½ΠΈΠ΅ΠΌ ΠΏΡΠΎΡΠ΅Π΄ΡΡΡ Π°ΡΡΠΈΠ½Π½ΠΎΠ³ΠΎ Π²ΡΠ΄Π΅Π»Π΅Π½ΠΈΡ ΠΈΠ· Π½Π΅Π³ΠΎ Π±Π΅Π»ΠΊΠΎΠ²-ΠΏΠ°ΡΡΠ½Π΅ΡΠΎΠ² Π΄Π»Ρ ΡΠ΅Π»Π΅Π²ΠΎΠ³ΠΎ Π±Π΅Π»ΠΊΠ° (Π±Π΅Π»ΠΊΠ°-Π½Π°ΠΆΠΈΠ²ΠΊΠΈ), ΠΈΠΌΠΌΠΎΠ±ΠΈΠ»ΠΈΠ·ΠΎΠ²Π°Π½Π½ΠΎΠ³ΠΎ Π½Π° ΠΈΠ½Π΅ΡΡΠ½ΠΎΠΌ ΡΠΎΡΠ±Π΅Π½ΡΠ΅ ΠΈΠ»ΠΈ ΠΏΠ°ΡΠ°ΠΌΠ°Π³Π½ΠΈΡΠ½ΡΡ
Π½Π°Π½ΠΎΡΠ°ΡΡΠΈΡΠ°Ρ
. Π¦ΠΈΠΊΠ» Π½Π°ΡΠΈΡ
ΠΏΡΠ΅Π΄ΡΠ΄ΡΡΠΈΡ
ΡΠ°Π±ΠΎΡ, ΠΏΠΎΡΠ²ΡΡΠ΅Π½Π½ΡΡ
ΠΏΡΡΠΌΠΎΠΌΡ ΠΌΠΎΠ»Π΅ΠΊΡΠ»ΡΡΠ½ΠΎΠΌΡ ΡΠΈΡΠΈΠ½Π³Ρ Ρ ΡΠΎΠΏΡΡΠΆΠ΅Π½ΠΈΠ΅ΠΌ Ρ
ΡΠΎΠΌΠ°ΡΠΎΠ³ΡΠ°ΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΈ ΠΌΠ°ΡΡ-ΡΠΏΠ΅ΠΊΡΡΠΎΠΌΠ΅ΡΡΠΈΡΠ΅ΡΠΊΠΈΡ
ΠΌΠ΅ΡΠΎΠ΄ΠΎΠ² ΠΈ ΡΠ΅Ρ
Π½ΠΎΠ»ΠΎΠ³ΠΈΠΈ ΠΏΠ°ΡΠ°ΠΌΠ°Π³Π½ΠΈΡΠ½ΡΡ
Π½Π°Π½ΠΎΡΠ°ΡΡΠΈΡ c ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΡΡΠ΄Π° Π±Π΅Π»ΠΊΠΎΠ² 18-ΠΎΠΉ Ρ
ΡΠΎΠΌΠΎΡΠΎΠΌΡ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ° ΠΈ ΡΠ°ΠΊΠΆΠ΅ Π΄ΡΡΠ³ΠΈΡ
Π±Π΅Π»ΠΊΠΎΠ² ΠΏΠΎΠΊΠ°Π·Π°Π», ΡΡΠΎ ΡΡΡΠ΅ΡΡΠ²ΡΡΡ, ΠΏΠΎ ΠΊΡΠ°ΠΉΠ½Π΅ ΠΌΠ΅ΡΠ΅, Π΄Π²Π΅ ΠΏΡΠΎΠ±Π»Π΅ΠΌΡ, Π²Π»ΠΈΡΡΡΠΈΠ΅ Π½Π° ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ½ΠΎΡΡΡ ΠΈ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΡ Π΄Π°Π½Π½ΠΎΠΉ ΠΏΡΠΎΡΠ΅Π΄ΡΡΡ: (i) ΠΈΠ·Π±ΡΡΠΎΡΠ½ΠΎΡΡΡ ΡΠΎΠ½ΠΎΠ²ΡΡ
Π±Π΅Π»ΠΊΠΎΠ² Π² ΡΠ»ΡΠ°ΡΠ°Ρ
Ρ Π°ΡΡΠΈΠ½Π½ΠΎΠ³ΠΎ ΡΠΎΡΠ±Π΅Π½ΡΠ°, ΠΎΠ±ΡΡΠ»ΠΎΠ²Π»Π΅Π½Π½Π°Ρ Π²ΡΠ΄Π΅Π»Π΅Π½ΠΈΠ΅ΠΌ ΠΌΡΠ»ΡΡΠΈΠ±Π΅Π»ΠΊΠΎΠ²ΡΡ
ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠΎΠ², ΠΌΠ΅ΡΠ΅Π½ΡΡ
ΠΏΡΡΠΌΡΠΌ ΠΏΠ°ΡΡΠ½Π΅ΡΠΎΠΌ, ΠΊΠΎΡΠΎΡΡΠΉ ΡΠ²ΡΠ·ΡΠ²Π°Π΅ΡΡΡ Ρ ΡΠ΅Π»Π΅Π²ΡΠΌ Π±Π΅Π»ΠΊΠΎΠΌ Π½Π° ΡΠΎΡΠ±Π΅Π½ΡΠ΅; (ii) Π½ΠΈΠ·ΠΊΠ°Ρ ΠΎΠ±ΠΎΠ³Π°ΡΠ΅Π½Π½ΠΎΡΡΡ ΡΠ»ΡΠ°ΡΠΎΠ² Π±Π΅Π»ΠΊΠ°ΠΌΠΈ-ΠΏΠ°ΡΡΠ½Π΅ΡΠ°ΠΌΠΈ ΡΠ΅Π»Π΅Π²ΠΎΠΉ Π³ΡΡΠΏΠΏΡ ΠΎΠ±ΡΡΠ»ΠΎΠ²Π»Π΅Π½Π½Π°Ρ ΡΠ΅ΠΌ, ΡΡΠΎ ΡΠ° ΠΈΠ»ΠΈ ΠΈΠ½Π°Ρ ΡΠ°ΡΡΡ ΠΏΡΡΠΌΡΡ
Π±Π΅Π»ΠΊΠΎΠ²-ΠΏΠ°ΡΡΠ½Π΅ΡΠΎΠ² Π² Π»ΠΈΠ·Π°ΡΠ΅ Π½Π°Ρ
ΠΎΠ΄ΠΈΡΡΡ Π² ΡΠΎΡΡΠ°Π²Π΅ ΡΡΠ°Π±ΠΈΠ»ΡΠ½ΡΡ
ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠΎΠ² Β«Π΄ΠΈΠΊΠΎΠ³ΠΎ ΡΠΈΠΏΠ°Β» Ρ ΡΠ°ΠΌΠΈΠΌ Π±Π΅Π»ΠΊΠΎΠΌ-Π½Π°ΠΆΠΈΠ²ΠΊΠΎΠΉ ΠΈ Π½Π΅ Π±ΡΠ΄Π΅Ρ Π² Π΄ΠΎΡΡΠ°ΡΠΎΡΠ½ΠΎΠΉ ΡΡΠ΅ΠΏΠ΅Π½ΠΈ Π²ΡΠ΄Π΅Π»Π΅Π½Π° ΠΈΠ· Π»ΠΈΠ·Π°ΡΠ°. ΠΠΎΡΡΠΎΠΌΡ Π΄Π»Ρ ΠΏΠΎΠ²ΡΡΠ΅Π½ΠΈΡ ΡΠΏΠ΅ΡΠΈΡΠΈΡΠ½ΠΎΡΡΠΈ ΠΈ ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ Π°ΡΡΠΈΠ½Π½ΠΎΠ³ΠΎ Π²ΡΠ΄Π΅Π»Π΅Π½ΠΈΡ Π±Π΅Π»ΠΊΠΎΠ²-ΠΏΠ°ΡΡΠ½Π΅ΡΠΎΠ² ΡΠ΅Π»Π΅Π²ΠΎΠ³ΠΎ Π±Π΅Π»ΠΊΠ° Π½Π°ΠΌΠΈ ΠΏΡΠ΅Π΄Π»ΠΎΠΆΠ΅Π½Π° ΠΌΠΎΠ΄ΠΈΡΠΈΠΊΠ°ΡΠΈΡ ΡΡΠ°Π½Π΄Π°ΡΡΠ½ΠΎΠΉ ΠΏΡΠΎΠ±ΠΎΠΏΠΎΠ΄Π³ΠΎΡΠΎΠ²ΠΊΠΈ, Π·Π°ΠΊΠ»ΡΡΠ°ΡΡΠ°ΡΡΡ Π² ΠΏΡΠ΅Π΄Π²Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎΠΉ Π΄ΠΈΡΡΠΎΡΠΈΠ°ΡΠΈΠΈ Π±Π΅Π»ΠΊΠΎΠ²ΡΡ
ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠΎΠ² Π»ΠΈΠ·Π°ΡΠ°. ΠΠΎΠ΄Π΅Π»ΡΠ½ΡΠ΅ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΡ ΠΏΠΎ Π²ΡΠ±ΠΎΡΡ ΡΠ΅Π³Π΅Π½Π΅ΡΠ°ΡΠΈΠΎΠ½Π½ΠΎΠ³ΠΎ ΡΠ°ΡΡΠ²ΠΎΡΠ°, ΠΎΡΠ΅Π½ΠΊΠ΅ ΡΡΠ°Π±ΠΈΠ»ΡΠ½ΠΎΡΡΠΈ ΠΈ ΡΠ²ΡΠ·ΡΠ²Π°ΡΡΠ΅ΠΉ ΡΠΏΠΎΡΠΎΠ±Π½ΠΎΡΡΠΈ Π±Π΅Π»ΠΊΠΎΠ² ΠΏΡΠΈ Π΅Π³ΠΎ Π²ΠΎΠ·Π΄Π΅ΠΉΡΡΠ²ΠΈΠΈ, Π° ΡΠ°ΠΊΠΆΠ΅ ΠΎΡΠ΅Π½ΠΊΠ° ΡΡΡΠ΅ΠΊΡΠΈΠ²Π½ΠΎΡΡΠΈ Π΄ΠΈΡΡΠΎΡΠΈΠ°ΡΠΈΠΈ ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠΎΠ² Π² Π»ΠΈΠ·Π°ΡΠ΅ Π±ΡΠ»ΠΈ Π²ΡΠΏΠΎΠ»Π½Π΅Π½Ρ ΠΏΠΎΠ΄ ΠΊΠΎΠ½ΡΡΠΎΠ»Π΅ΠΌ ΠΎΠΏΡΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ Π±ΠΈΠΎΡΠ΅Π½ΡΠΎΡΠ° Biacore 3000 (Β«GE HealthcareΒ», Π‘Π¨Π) Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ Π»ΠΈΠ·Π°ΡΠ° ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ ΠΊΡΠ»ΡΡΡΡΡ Π³Π΅ΠΏΠ°ΡΠΎΠΊΠ°ΡΡΠΈΠ½ΠΎΠΌΡ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ° (HepG2) ΠΈ ΡΠ΅ΠΊΠΎΠΌΠ±ΠΈΠ½Π°Π½ΡΠ½ΡΡ
ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠΎΠ² Π±Π΅Π»ΠΊΠΎΠ², ΠΊΠΎΠ΄ΠΈΡΡΠ΅ΠΌΡΡ
Π³Π΅Π½Π°ΠΌΠΈ 18-ΠΎΠΉ Ρ
ΡΠΎΠΌΠΎΡΠΎΠΌΡ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ°, ΠΠΎΠΊΠ°Π·Π°Π½ΠΎ, ΡΡΠΎ ΠΊΠΈΡΠ»ΠΎΡΠ½Π°Ρ ΠΎΠ±ΡΠ°Π±ΠΎΡΠΊΠ° ΡΠ°Π·Π±Π°Π²Π»Π΅Π½Π½ΠΎΠ³ΠΎ Π² 20 ΡΠ°Π· Π»ΠΈΠ·Π°ΡΠ° Ρ ΠΊΡΠ°ΡΠΊΠΎΠ²ΡΠ΅ΠΌΠ΅Π½Π½ΠΎΠΉ ΡΠΊΡΠΏΠΎΠ·ΠΈΡΠΈΠ΅ΠΉ Π² ΡΠ΅ΡΠ΅Π½ΠΈΠ΅ 1 ΠΌΠΈΠ½ Π½Π° Π»ΡΠ΄Ρ ΠΈ Ρ ΠΏΠΎΡΠ»Π΅Π΄ΡΡΡΠ΅ΠΉ Π½Π΅ΠΉΡΡΠ°Π»ΠΈΠ·Π°ΡΠΈΠ΅ΠΉ (Ρ ΡΠ 2.0 Π΄ΠΎ ΡΠ 7.4) ΠΏΡΠΈΠ²ΠΎΠ΄ΠΈΠ»Π° ΠΊ ΠΌΠ°ΠΊΡΠΈΠΌΠ°Π»ΡΠ½ΠΎΠΉ Π΄ΠΈΡΡΠΎΡΠΈΠ°ΡΠΈΠΈ Π±Π΅Π»ΠΊΠΎΠ²ΡΡ
ΠΊΠΎΠΌΠΏΠ»Π΅ΠΊΡΠΎΠ² Π»ΠΈΠ·Π°ΡΠ°, Π½Π΅ ΠΎΠΊΠ°Π·ΡΠ²Π°Ρ ΡΡΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠ³ΠΎ Π½Π΅Π³Π°ΡΠΈΠ²Π½ΠΎΠ³ΠΎ Π²Π»ΠΈΡΠ½ΠΈΡ Π½Π° ΡΠ΅ΡΡΠΈΡΡΠ΅ΠΌΡΠ΅ Π±Π΅Π»ΠΎΠΊ- Π±Π΅Π»ΠΊΠΎΠ²ΡΠ΅ Π²Π·Π°ΠΈΠΌΠΎΠ΄Π΅ΠΉΡΡΠ²ΠΈΡ
The main factors of poverty in modern Russian society
Background. In modern Russian society, the problem of poverty is relevant,
which necessitates the study of the main factors affecting the level of populationβs poverty.
It is important to take into account the approaches used to measure the level of the poor
society, what factors and causes influence the risk of falling into poverty, including in the
context of regional characteristics. The purpose of the study is to analyze the main factors
affecting the level of populationβs poverty in modern Russian society. Materials and methods.
Systematic and structural-functional approaches were used as methodological bases.
Based on the results of the authorβs research among the population of Arkhangelsk region
in 2022, a sociological analysis of the standard of living of the population in the region was
carried out, the main factors affecting the risk of falling into poverty, supplemented by official
data from Rosstat on the standard of living in Arkhangelsk region and the poorest regions
of Russia. Results. Based on the data of authorβs research, the main factors affecting
the level of poverty in Arkhangelsk region were analyzed. The main factors that have a
significant impact on the risk of people falling into poverty include: economic (low wages,
debt obligations); demographic (family composition, high degree of dependent burden);
educational and qualification (availability of education and high qualifications); medical
(health system availability and poor health). The use of official data of the Federal State
Statistics Service made it possible to identify the level of poverty in society and certain
regions of Russia, determine the nature of poverty in modern conditions, and establish the
risks of entering the poor. Conclusions. The study showed that poverty in modern conditions
in some cases is stagnant. The population of Arkhangelsk region has no difficulties in
satisfying the primary needs, but the purchase of durable goods causes material difficulties
in purchasing, which allows us to talk about the presence of a difficult financial situation in
families. The impact of the economic factor on the level of poverty in the region is determined
by the low level of wages, an increase in family spending and a high level of debt
burden. The demographic factor of poverty has revealed an increased dependent burden in
low-income families. The educational and qualification factor drew the attention of researchers
to a large number of working poor in Arkhangelsk region. Poor availability of
health services can also hinder overcoming low financial situation
ΠΠ΄Π΅Π½ΡΠΈΡΠΈΠΊΠ°ΡΠΈΡ ΡΠΈΠ±ΠΎΡΠΎΠΌΠ½ΡΡ ΡΡΡΠΏΡΠΈΠ½ΡΠΎΠ² Π½Π° ΡΠ»Π΅ΠΊΡΡΠΎΡΠΎΡΠ΅ΡΠΈΡΠ΅ΡΠΊΠΎΠΌ Π³Π΅Π»Π΅ ΠΏΡΠΈ ΠΏΡΠΎΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΠΈ ΡΡΠ°Π½ΡΠ»Π°ΡΠΎΠΌΠ°: ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ ΠΠΠ-ΠΌΠ°ΡΠΊΡΡΠΎΠ²
The commercial DNA ladder was tested as a substitute for RNA size standards to identify ribosomal footprints (RNA fragments of about 30 nucleotides long) on an electrophoretic polyacrylamide gel for the purposes of translatome profiling. It has been found that 25 and 35 nucleotides long synthetic RNA oligonucleotides do migrate slower than the synthetic DNA oligonucleotides of the matching length and sequences and their positions on the gel coincide with those of 30 and 40 nucleotides long DNA oligonucleotides, correspondingly, of the commercial IDT 20/100 DNA oligo length standards. By using this DNA ladder and RNA isolated from the preparation enriched in ribosomes (obtained by fractionating on MicroSpin S-400 columns the HepG2 cell lysate treated with RNase I), the position of a band of putative ribosomal footprints can be identified on a gel that has been verified by measuring in an RNA-seq experiment the length of RNA fragments extracted from the band.ΠΠΎΠΌΠΌΠ΅ΡΡΠ΅ΡΠΊΠΈΠ΅ ΠΠΠ-ΠΌΠ°ΡΠΊΡΡΡ ΡΠ΅ΡΡΠΈΡΠΎΠ²Π°Π»ΠΈ ΠΊΠ°ΠΊ Π·Π°ΠΌΠ΅Π½Ρ Π ΠΠ-ΡΡΠ°Π½Π΄Π°ΡΡΠΎΠ² Π΄Π»ΠΈΠ½Ρ ΠΌΠΎΠ»Π΅ΠΊΡΠ»Ρ Π΄Π»Ρ ΠΈΠ΄Π΅Π½ΡΠΈΡΠΈΠΊΠ°ΡΠΈΠΈ ΡΠΈΠ±ΠΎΡΠΎΠΌΠ½ΡΡ
ΡΡΡΠΏΡΠΈΠ½ΡΠΎΠ² (ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² Π ΠΠ Π΄Π»ΠΈΠ½ΠΎΠΉ ΠΎΠΊΠΎΠ»ΠΎ 30 Π½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄ΠΎΠ²) Π½Π° ΡΠ»Π΅ΠΊΡΡΠΎΡΠΎΡΠ΅ΡΠΈΡΠ΅ΡΠΊΠΎΠΌ ΠΏΠΎΠ»ΠΈΠ°ΠΊΡΠΈΠ»Π°ΠΌΠΈΠ΄Π½ΠΎΠΌ Π³Π΅Π»Π΅ ΠΏΡΠΈ ΠΏΡΠΎΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΠΈ ΡΡΠ°Π½ΡΠ»Π°ΡΠΎΠΌΠ°. ΠΡΠ»ΠΎ ΠΎΠ±Π½Π°ΡΡΠΆΠ΅Π½ΠΎ, ΡΡΠΎ ΡΠΈΠ½ΡΠ΅ΡΠΈΡΠ΅ΡΠΊΠΈΠ΅ Π ΠΠ-ΠΎΠ»ΠΈΠ³ΠΎΠ½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄Ρ Π΄Π»ΠΈΠ½ΠΎΠΉ 25 ΠΈ 35 Π½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄ΠΎΠ² ΠΌΠΈΠ³ΡΠΈΡΡΡΡ ΠΌΠ΅Π΄Π»Π΅Π½Π½Π΅Π΅, ΡΠ΅ΠΌ ΡΠΈΠ½ΡΠ΅ΡΠΈΡΠ΅ΡΠΊΠΈΠ΅ ΠΠΠ-ΠΎΠ»ΠΈΠ³ΠΎΠ½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄Ρ ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²ΡΡΡΠ΅ΠΉ Π΄Π»ΠΈΠ½Ρ ΠΈ ΠΏΠΎΡΠ»Π΅Π΄ΠΎΠ²Π°ΡΠ΅Π»ΡΠ½ΠΎΡΡΠ΅ΠΉ, ΠΈ ΠΈΡ
ΠΏΠΎΠ»ΠΎΠΆΠ΅Π½ΠΈΠ΅ Π½Π° Π³Π΅Π»Π΅ ΡΠΎΠ²ΠΏΠ°Π΄Π°Π΅Ρ Ρ ΠΏΠΎΠ»ΠΎΠΆΠ΅Π½ΠΈΠ΅ΠΌ ΠΠΠ- ΠΎΠ»ΠΈΠ³ΠΎΠ½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄ΠΎΠ² Π΄Π»ΠΈΠ½ΠΎΠΉ 30 ΠΈ 40 Π½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄ΠΎΠ² ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²Π΅Π½Π½ΠΎ ΠΈΠ· ΠΊΠΎΠΌΠΌΠ΅ΡΡΠ΅ΡΠΊΠΎΠ³ΠΎ Π½Π°Π±ΠΎΡΠ° ΡΡΠ°Π½Π΄Π°ΡΡΠΎΠ² Π΄Π»ΠΈΠ½ ΠΠΠ-ΠΎΠ»ΠΈΠ³ΠΎΠ½ΡΠΊΠ»Π΅ΠΎΡΠΈΠ΄ΠΎΠ² Β«IDT 20/100 DNA oligo length standardsΒ». ΠΡΠΏΠΎΠ»ΡΠ·ΡΡ Π΄Π°Π½Π½ΡΠΉ Π½Π°Π±ΠΎΡ ΠΈ Π ΠΠ, Π²ΡΠ΄Π΅Π»Π΅Π½Π½ΡΡ ΠΈΠ· ΠΏΡΠ΅ΠΏΠ°ΡΠ°ΡΠ°, ΠΎΠ±ΠΎΠ³Π°ΡΠ΅Π½Π½ΠΎΠ³ΠΎ ΡΠΈΠ±ΠΎΡΠΎΠΌΠ°ΠΌΠΈ (ΠΏΠΎΠ»ΡΡΠ΅Π½Π½ΠΎΠ³ΠΎ ΡΡΠ°ΠΊΡΠΈΠΎΠ½ΠΈΡΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ Π½Π° ΠΊΠΎΠ»ΠΎΠ½ΠΊΠ°Ρ
MicroSpin S-400 ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠ³ΠΎ Π»ΠΈΠ·Π°ΡΠ° HepG2, ΠΎΠ±ΡΠ°Π±ΠΎΡΠ°Π½Π½ΠΎΠ³ΠΎ Π ΠΠΠ°Π·ΠΎΠΉ I), ΠΌΠΎΠΆΠ½ΠΎ ΠΈΠ΄Π΅Π½ΡΠΈΡΠΈΡΠΈΡΠΎΠ²Π°ΡΡ ΠΏΠΎΠ»ΠΎΠΆΠ΅Π½ΠΈΠ΅ ΠΏΠΎΠ»ΠΎΡΡ ΠΏΡΠ΅Π΄ΠΏΠΎΠ»Π°Π³Π°Π΅ΠΌΡΡ
ΡΠΈΠ±ΠΎΡΠΎΠΌΠ½ΡΡ
ΡΡΡΠΏΡΠΈΠ½ΡΠΎΠ² Π½Π° Π³Π΅Π»Π΅, ΡΡΠΎ Π±ΡΠ»ΠΎ ΠΏΠΎΠ΄ΡΠ²Π΅ΡΠΆΠ΄Π΅Π½ΠΎ ΠΈΠ·ΠΌΠ΅ΡΠ΅Π½ΠΈΠ΅ΠΌ Π΄Π»ΠΈΠ½Ρ ΡΡΠ°Π³ΠΌΠ΅Π½ΡΠΎΠ² Π ΠΠ, Π²ΡΠ΄Π΅Π»Π΅Π½Π½ΡΡ
ΠΈΠ· ΠΏΠΎΠ»ΠΎΡΡ, ΠΏΡΠΈ ΠΏΡΠΎΠ²Π΅Π΄Π΅Π½ΠΈΠΈ ΠΈΡ
ΡΠ΅ΠΊΠ²Π΅Π½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ RNA-seq
Osteogenic potential of multipotent mesenchymal stromal cells from human exfoliated deciduous teeth before and after cryopreservation
The use of multipotent mesenchymal stromal cellsfrom human exfoliated deciduous teeth (SHED) to stimulate bone regeneration requires data on the influence of cryopreservation on the osteogenic differentiation capacity of these cells. SHED were subjected to cryopreservation. Before freezing and after thawing, cell cultures were exposed osteogenic differentiation with vitamin D3 or dexametasone and assessed for expression of the osteogenic markers osteocalcin, alkaline phosphatase, BMP-2 and RunX2 using real-Time qPCR. Extracellular matrix (ECM) mineralization was evaluated by Alizarin red staining. Supplementation of osteogenic medium with vitamin D3 increased the expression of the osteogenic markers osteocalcin, alkaline phosphatase, BMP-2 and RunX2 as well as promoted an increase in the synthesis and mineralization of ECM in the cells both before and after cryopreservation. In the presence of vitamin D3 gene expression of alkaline phosphatase, BMP-2 and RunX2 after cryopreservation was higher than before freezing. Gene expression of osteocalcin, BMP2 RunX2 in osteogenic medium with vitamin D3was higher compared with dexamethasone for 14 days differentiation both before or after cryopreservation. The maintenance of SHED osteogenic differentiation potential after long-Term cryopreservation provides a basis for banking of these cellsfor further auto- or allotransplantation
Osteogenic potential of multipotent mesenchymal stromal cells from human exfoliated deciduous teeth before and after cryopreservation
The use of multipotent mesenchymal stromal cellsfrom human exfoliated deciduous teeth (SHED) to stimulate bone regeneration requires data on the influence of cryopreservation on the osteogenic differentiation capacity of these cells. SHED were subjected to cryopreservation. Before freezing and after thawing, cell cultures were exposed osteogenic differentiation with vitamin D3 or dexametasone and assessed for expression of the osteogenic markers osteocalcin, alkaline phosphatase, BMP-2 and RunX2 using real-Time qPCR. Extracellular matrix (ECM) mineralization was evaluated by Alizarin red staining. Supplementation of osteogenic medium with vitamin D3 increased the expression of the osteogenic markers osteocalcin, alkaline phosphatase, BMP-2 and RunX2 as well as promoted an increase in the synthesis and mineralization of ECM in the cells both before and after cryopreservation. In the presence of vitamin D3 gene expression of alkaline phosphatase, BMP-2 and RunX2 after cryopreservation was higher than before freezing. Gene expression of osteocalcin, BMP2 RunX2 in osteogenic medium with vitamin D3was higher compared with dexamethasone for 14 days differentiation both before or after cryopreservation. The maintenance of SHED osteogenic differentiation potential after long-Term cryopreservation provides a basis for banking of these cellsfor further auto- or allotransplantation
ΠΠ΅Π½Ρ Β«ΡΡΠ°Ρ Π°Π½ΠΎΠ²ΡΡΒ» 18 Ρ ΡΠΎΠΌΠΎΡΠΎΠΌΡ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ°, ΠΎΡΡΡΡΡΡΠ²ΡΡΡΠΈΠ΅ Π±Π΅Π»ΠΊΠΈ ΠΈ Π½Π΅ ΠΎΡ Π°ΡΠ°ΠΊΡΠ΅ΡΠΈΠ·ΠΎΠ²Π°Π½Π½ΡΠ΅ Π±Π΅Π»ΠΊΠΈ Π² ΡΠΊΠ°Π½ΠΈ ΠΏΠ΅ΡΠ΅Π½ΠΈ ΠΈ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ Π»ΠΈΠ½ΠΈΠΈ HepG2
Missing (MP) and functionally uncharacterized proteins (uPE1) comprise less than 5% of the total number of proteins encoded by human Chr18 genes. Within half a year, since the January 2020 version of NextProt, the number of entries in the MP+uPE1 datasets changed, mainly due to the achievements of antibody-based proteomics. Assuming that the proteome is closely related to the transcriptome scaffold, quantitative PCR, Illumina HiSeq, and Oxford Nanopore Technology were applied to characterize the liver samples of three male donors in comparison with the HepG2 cell line. The data mining of the Expression Atlas (EMBL-EBI) and the profiling of biopsy samples by using orthogonal methods of transcriptome analysis have shown that in HepG2 cells and the liver, the genes encoding functionally uncharacterized proteins (uPE1) are expressed as low as for the missing proteins (less than 1 copy per cell), except the selected cases of HSBP1L1, TMEM241, C18orf21, and KLHL14. The initial expectation that uPE1 genes might be expressed at higher levels than MP genes, was compromised by severe discrepancies in our semi-quantitative gene expression data and in public databanks. Such discrepancy forced us to revisit the transcriptome of Chr18, the target of the Russian C-HPP Consortium. Tanglegram of highly expressed genes and further correlation analysis have shown the severe dependencies on the mRNA extraction method and the analytical platform. Targeted gene expression analysis by quantitative PCR (qPCR) and high-throughput transcriptome profiling (Illumina HiSeq and ONT MinION) for the same set of samples from normal liver tissue and HepG2 cells revealed the detectable expression of 250+ (92%) protein-coding genes of Chr18 (at least one method). The expression of slightly more than 50% protein-coding genes was detected simultaneously by all three methods. Correlation analysis of the gene expression profiles showed that the grouping of the datasets depended almost equally on both the type of biological material and the experimental method, particularly cDNA/mRNA isolation and library preparation.ΠΡΡΡΡΡΡΠ²ΡΡΡΠΈΠ΅ Π±Π΅Π»ΠΊΠΈ ΠΈ ΡΡΠ½ΠΊΡΠΈΠΎΠ½Π°Π»ΡΠ½ΠΎ Π½Π΅ ΠΎΡ
Π°ΡΠ°ΠΊΡΠ΅ΡΠΈΠ·ΠΎΠ²Π°Π½Π½ΡΠ΅ Π±Π΅Π»ΠΊΠΈ (Π² Π°Π½Π³Π»ΠΎΡΠ·ΡΡΠ½ΠΎΠΉ Π»ΠΈΡΠ΅ΡΠ°ΡΡΡΠ΅ ΠΎΠ±ΠΎΠ·Π½Π°ΡΠ΅Π½Π½ΡΠ΅ ΠΊΠ°ΠΊ missing (MP) ΠΈ functionally uncharacterized proteins (uPE1), ΡΠΎΠΎΡΠ²Π΅ΡΡΡΠ²Π΅Π½Π½ΠΎ) ΡΠΎΡΡΠ°Π²Π»ΡΡΡ ΠΌΠ΅Π½Π΅Π΅ 5% ΠΎΡ ΠΎΠ±ΡΠ΅Π³ΠΎ ΡΠΈΡΠ»Π° Π±Π΅Π»ΠΊΠΎΠ², ΠΊΠΎΠ΄ΠΈΡΡΠ΅ΠΌΡΡ
Π³Π΅Π½Π°ΠΌΠΈ 18 Ρ
ΡΠΎΠΌΠΎΡΠΎΠΌΡ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ°. Π ΡΠ΅ΡΠ΅Π½ΠΈΠ΅ ΠΏΠΎΠ»ΡΠ³ΠΎΠ΄Π°, Π½Π°ΡΠΈΠ½Π°Ρ Ρ ΡΠ½Π²Π°ΡΡ 2020 Π³ΠΎΠ΄Π°, Π² Π²Π΅ΡΡΠΈΠΈ NextProt Π²ΡΡΠΎΡΠ»ΠΎ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²ΠΎ Π·Π°ΠΏΠΈΡΠ΅ΠΉ Π² Π½Π°Π±ΠΎΡΠ°Ρ
Π΄Π°Π½Π½ΡΡ
MP+uPE1. ΠΠΎΠ΄ΠΎΠ±Π½ΡΠ΅ ΠΈΠ·ΠΌΠ΅Π½Π΅Π½ΠΈΡ ΠΎΠ±ΡΡΠ»ΠΎΠ²Π»Π΅Π½Ρ ΠΏΡΠ΅ΠΈΠΌΡΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎ Π΄ΠΎΡΡΠΈΠΆΠ΅Π½ΠΈΡΠΌΠΈ ΠΏΡΠΎΡΠ΅ΠΎΠΌΠΈΠΊΠΈ Π½Π° ΠΎΡΠ½ΠΎΠ²Π΅ Π°Π½ΡΠΈΡΠ΅Π». Π Π΄Π°Π½Π½ΠΎΠΉ ΡΠ°Π±ΠΎΡΠ΅ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½Π°Ρ ΠΠ¦Π , ΡΠ΅Ρ
Π½ΠΎΠ»ΠΎΠ³ΠΈΠΈ ΡΠ΅ΠΊΠ²Π΅Π½ΠΈΡΠΎΠ²Π°Π½ΠΈΡ Illumina HiSeq ΠΈ Oxford Nanopore Technologies Π±ΡΠ»ΠΈ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½Ρ Π΄Π»Ρ ΡΡΠ°Π²Π½ΠΈΡΠ΅Π»ΡΠ½ΠΎΠ³ΠΎ Π°Π½Π°Π»ΠΈΠ·Π° ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΎΠΌΠ½ΠΎΠ³ΠΎ ΠΏΡΠΎΡΠΈΠ»Ρ ΠΎΠ±ΡΠ°Π·ΡΠΎΠ² ΠΏΠ΅ΡΠ΅Π½ΠΈ ΡΡΠ΅Ρ
Π΄ΠΎΠ½ΠΎΡΠΎΠ² ΠΌΡΠΆΡΠΊΠΎΠ³ΠΎ ΠΏΠΎΠ»Π° ΠΈ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ Π»ΠΈΠ½ΠΈΠΈ HepG2. ΠΠ½Π°Π»ΠΈΠ· Π΄Π°Π½Π½ΡΡ
Π°ΡΠ»Π°ΡΠ° ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ (Expression Atlas, EMBL-EBI) ΠΈ ΠΏΠΎΠ»ΡΡΠ΅Π½Π½ΡΡ
ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΠΎΠ² ΠΏΠΎ Π±ΠΈΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΈΠΌ ΠΎΠ±ΡΠ°Π·ΡΠ°ΠΌ Ρ ΠΈΡΠΏΠΎΠ»ΡΠ·ΠΎΠ²Π°Π½ΠΈΠ΅ΠΌ ΠΎΡΡΠΎΠ³ΠΎΠ½Π°Π»ΡΠ½ΡΡ
ΠΌΠ΅ΡΠΎΠ΄ΠΎΠ² Π°Π½Π°Π»ΠΈΠ·Π° ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΎΠΌΠ° ΠΏΠΎΠΊΠ°Π·Π°Π», ΡΡΠΎ Π² ΠΊΠ»Π΅ΡΠΊΠ°Ρ
ΠΏΠ΅ΡΠ΅Π½ΠΈ ΠΈ HepG2 ΡΡΠΎΠ²Π΅Π½Ρ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ Π³Π΅Π½ΠΎΠ², ΠΊΠΎΠ΄ΠΈΡΡΡΡΠΈΡ
ΡΡΠ½ΠΊΡΠΈΠΎΠ½Π°Π»ΡΠ½ΠΎ Π½Π΅ ΠΎΡ
Π°ΡΠ°ΠΊΡΠ΅ΡΠΈΠ·ΠΎΠ²Π°Π½Π½ΡΠ΅ Π±Π΅Π»ΠΊΠΈ (uPE1), Π½Π°Ρ
ΠΎΠ΄ΠΈΡΡΡ Π½Π° ΡΠ°ΠΊΠΎΠΌ ΠΆΠ΅ Π½ΠΈΠ·ΠΊΠΎΠΌ ΡΡΠΎΠ²Π½Π΅, ΠΊΠ°ΠΊ ΠΈ Π² ΡΠ»ΡΡΠ°Π΅ Π³Π΅Π½ΠΎΠ² MP (Π² ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅ ΠΌΠ΅Π½Π΅Π΅ 1 ΠΊΠΎΠΏΠΈΠΈ Π½Π° ΠΊΠ»Π΅ΡΠΊΡ). ΠΡΠΊΠ»ΡΡΠ΅Π½ΠΈΠ΅ ΡΠΎΡΡΠ°Π²ΠΈΠ»ΠΈ Π½Π΅ΡΠΊΠΎΠ»ΡΠΊΠΎ Π³Π΅Π½ΠΎΠ²: HSBP1L1, TMEM241, C18orf21 ΠΈ KLHL14. Π‘ΠΎΠ³Π»Π°ΡΠ½ΠΎ ΡΡΡΠ΅ΡΡΠ²Π΅Π½Π½ΡΠΌ ΡΠ°ΡΡ
ΠΎΠΆΠ΄Π΅Π½ΠΈΡΠΌ Π² ΡΠ°Π½Π΅Π΅ ΠΏΠΎΠ»ΡΡΠ΅Π½Π½ΡΡ
ΠΏΠΎΠ»ΡΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΡΡ
Π΄Π°Π½Π½ΡΡ
ΠΏΠΎ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ Π³Π΅Π½ΠΎΠ² ΠΈ Π΄Π°Π½Π½ΡΠΌ Π² ΠΎΡΠΊΡΡΡΡΡ
Π±Π°Π·Π°Ρ
Π΄Π°Π½Π½ΡΡ
, ΠΈΠ·Π½Π°ΡΠ°Π»ΡΠ½ΠΎ ΠΏΡΠ΅Π΄ΠΏΠΎΠ»Π°Π³Π°Π»ΠΎΡΡ, ΡΡΠΎ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΡ Π³Π΅Π½ΠΎΠ² uPE1 ΠΌΠΎΠΆΠ΅Ρ Π±ΡΡΡ Π²ΡΡΠ΅, ΡΠ΅ΠΌ Π³Π΅Π½ΠΎΠ² MP. ΠΠΎΠ΄ΠΎΠ±Π½ΠΎΠ΅ ΡΠ°ΡΡ
ΠΎΠΆΠ΄Π΅Π½ΠΈΠ΅ ΠΏΠΎΠ±ΡΠ΄ΠΈΠ»ΠΎ ΠΎΠ±ΡΠ°ΡΠΈΡΡΡΡ ΠΊ ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΎΠΌΡ 18 Ρ
ΡΠΎΠΌΠΎΡΠΎΠΌΡ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ°, ΡΠ²Π»ΡΡΡΠ΅ΠΉΡΡ ΡΠ΅Π»Π΅Π²ΠΎΠΉ Π΄Π»Ρ Π ΠΎΡΡΠΈΠΈ Π² ΠΏΡΠΎΠ΅ΠΊΡΠ΅ Β«ΠΡΠΎΡΠ΅ΠΎΠΌ ΡΠ΅Π»ΠΎΠ²Π΅ΠΊΠ°Β». ΠΠΎΠ»ΡΡΠ΅Π½Π½ΡΠ΅ ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΡ ΠΎ Π½Π°ΠΈΠ±ΠΎΠ»Π΅Π΅ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΡΡΠ΅ΠΌΡΡ
Π³Π΅Π½Π°Ρ
ΠΈ Π΄Π°Π»ΡΠ½Π΅ΠΉΡΠΈΠΉ ΠΊΠΎΡΡΠ΅Π»ΡΡΠΈΠΎΠ½Π½ΡΠΉ Π°Π½Π°Π»ΠΈΠ· ΠΏΠΎΠΊΠ°Π·Π°Π» ΡΡΡΠ΅ΡΡΠ²ΠΎΠ²Π°Π½ΠΈΠ΅ Π·Π°Π²ΠΈΡΠΈΠΌΠΎΡΡΠΈ ΠΎΡ ΠΌΠ΅ΡΠΎΠ΄Π° ΡΠΊΡΡΡΠ°ΠΊΡΠΈΠΈ ΠΌΠ ΠΠ ΠΈ Π°Π½Π°Π»ΠΈΡΠΈΡΠ΅ΡΠΊΠΎΠΉ ΠΏΠ»Π°ΡΡΠΎΡΠΌΡ. ΠΠ½Π°Π»ΠΈΠ· ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ ΡΠ΅Π»Π΅Π²ΡΡ
Π³Π΅Π½ΠΎΠ² 18 Ρ
ΡΠΎΠΌΠΎΡΠΎΠΌΡ Ρ ΠΏΡΠΈΠΌΠ΅Π½Π΅Π½ΠΈΠ΅ΠΌ ΠΊΠΎΠ»ΠΈΡΠ΅ΡΡΠ²Π΅Π½Π½ΠΎΠΉ ΠΠ¦Π (qPCR) ΠΈ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠ² Π²ΡΡΠΎΠΊΠΎΠΏΡΠΎΠΈΠ·Π²ΠΎΠ΄ΠΈΡΠ΅Π»ΡΠ½ΠΎΠ³ΠΎ ΠΏΡΠΎΡΠΈΠ»ΠΈΡΠΎΠ²Π°Π½ΠΈΡ ΡΡΠ°Π½ΡΠΊΡΠΈΠΏΡΠΎΠΌΠ° (Illumina HiSeq ΠΈ ONT MinION) Π΄Π»Ρ ΠΎΠ΄ΠΈΠ½Π°ΠΊΠΎΠ²ΡΡ
Π½Π°Π±ΠΎΡΠΎΠ² ΠΎΠ±ΡΠ°Π·ΡΠΎΠ² Π½ΠΎΡΠΌΠ°Π»ΡΠ½ΠΎΠΉ ΡΠΊΠ°Π½ΠΈ ΠΏΠ΅ΡΠ΅Π½ΠΈ ΠΈ ΠΊΠ»Π΅ΡΠΎΡΠ½ΠΎΠΉ Π»ΠΈΠ½ΠΈΠΈ HepG2 Π²ΡΡΠ²ΠΈΠ» Π±ΠΎΠ»Π΅Π΅ 250 (92%) Π±Π΅Π»ΠΎΠΊ-ΠΊΠΎΠ΄ΠΈΡΡΡΡΠΈΡ
Π³Π΅Π½ΠΎΠ², Π΄Π΅ΡΠ΅ΠΊΡΠΈΡΡΠ΅ΠΌΡΡ
Ρ
ΠΎΡΡ Π±Ρ ΠΎΠ΄Π½ΠΈΠΌ ΠΌΠ΅ΡΠΎΠ΄ΠΎΠΌ. ΠΠΊΡΠΏΡΠ΅ΡΡΠΈΡ Π±ΠΎΠ»Π΅Π΅ ΡΠ΅ΠΌ 50% Π±Π΅Π»ΠΎΠΊ-ΠΊΠΎΠ΄ΠΈΡΡΡΡΠΈΡ
Π³Π΅Π½ΠΎΠ² Π±ΡΠ»Π° Π΄Π΅ΡΠ΅ΠΊΡΠΈΡΠΎΠ²Π°Π½Π° Π²ΡΠ΅ΠΌΠΈ ΡΡΠ΅ΠΌΡ ΠΌΠ΅ΡΠΎΠ΄Π°ΠΌΠΈ. ΠΠΎΡΡΠ΅Π»ΡΡΠΈΠΎΠ½Π½ΡΠΉ Π°Π½Π°Π»ΠΈΠ· ΠΏΡΠΎΡΠΈΠ»Π΅ΠΉ ΡΠΊΡΠΏΡΠ΅ΡΡΠΈΠΈ Π³Π΅Π½ΠΎΠ² ΠΏΠΎΠΊΠ°Π·Π°Π», ΡΡΠΎ ΡΠ΅Π·ΡΠ»ΡΡΠ°ΡΡ Β«Π³ΡΡΠΏΠΏΠΈΡΡΡΡΡΡΒ» Π² Π·Π°Π²ΠΈΡΠΈΠΌΠΎΡΡΠΈ ΠΎΡ ΡΠΈΠΏΠ° Π±ΠΈΠΎΠ»ΠΎΠ³ΠΈΡΠ΅ΡΠΊΠΎΠ³ΠΎ ΠΌΠ°ΡΠ΅ΡΠΈΠ°Π»Π° ΠΈ ΡΠΊΡΠΏΠ΅ΡΠΈΠΌΠ΅Π½ΡΠ°Π»ΡΠ½ΡΡ
ΠΌΠ΅ΡΠΎΠ΄ΠΎΠ², Π² ΡΠ°ΡΡΠ½ΠΎΡΡΠΈ ΠΎΡ ΡΠΏΠΎΡΠΎΠ±Π° ΠΏΠΎΠ΄Π³ΠΎΡΠΎΠ²ΠΊΠΈ Π±ΠΈΠ±Π»ΠΈΠΎΡΠ΅ΠΊΠΈ (Π²ΡΠ΄Π΅Π»Π΅Π½ΠΈΡ ΠΊΠΠΠ, ΠΌΠ ΠΠ). ΠΠ°Π²ΠΈΡΠΈΠΌΠΎΡΡΡ ΠΎΡ Π²ΡΠ±ΠΎΡΠ° ΡΠΏΠΎΡΠΎΠ±Π° Π±ΠΈΠΎΠΈΠ½ΡΠΎΡΠΌΠ°ΡΠΈΡΠ΅ΡΠΊΠΎΠΉ ΠΎΠ±ΡΠ°Π±ΠΎΡΠΊΠΈ Π±ΡΠ»Π° ΠΎΡΠΌΠ΅ΡΠ΅Π½Π° Π² Π·Π½Π°ΡΠΈΡΠ΅Π»ΡΠ½ΠΎ ΠΌΠ΅Π½ΡΡΠ΅ΠΉ ΡΡΠ΅ΠΏΠ΅Π½ΠΈ