CN107541558B - Method for detecting capability of assembling chromatin structure on specific site of DNA sequence - Google Patents
Method for detecting capability of assembling chromatin structure on specific site of DNA sequence Download PDFInfo
- Publication number
- CN107541558B CN107541558B CN201710903858.2A CN201710903858A CN107541558B CN 107541558 B CN107541558 B CN 107541558B CN 201710903858 A CN201710903858 A CN 201710903858A CN 107541558 B CN107541558 B CN 107541558B
- Authority
- CN
- China
- Prior art keywords
- chromatin structure
- dna sequence
- specific site
- band
- assembling
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The application relates to the field of molecular biology, in particular to a method for detecting the capability of assembling a chromatin structure at a specific site of a DNA sequence, which is used for solving the problem that the prior art cannot detect the occupancy rate of nucleosomes at fixed points to determine the assembly capability of the chromatin structure. The method mainly comprises the steps of assembling DNA sequences containing specific restriction enzyme sites into a chromatin structure, carrying out enzyme digestion by using the restriction enzyme, removing various proteins, carrying out electrophoresis detection, quantitatively calculating the occupancy rate of nucleosomes of the sites and the DNA sequences nearby the sites, and judging the local chromatin structure nearby the detected sites. Furthermore, the efficiency of assembling the chromatin structure at a specific site on the DNA sequence and the assembly capacity of the nucleosomes are analyzed, and the detection method has the advantages of simple and convenient realization process and high specificity and accuracy.
Description
Technical Field
The application relates to the field of molecular biology, in particular to a method for detecting the capability of assembling a chromatin structure at a specific site of a DNA sequence.
Background
The nucleosome is a basic structural unit of eucaryon chromatin, two copies of each of four histones H2A, H2B, H3 and H4 form a histone octamer, and a DNA sequence of about 147bp is wound on the histone octamer in a left-hand spiral mode for 1.7 circles to form a nucleosome core particle. The core particles are connected by 20-80bp DNA, and are further compressed and packaged into a 30nm chromatin secondary structure under the action of histone H1.
As a basic structural unit of eucaryon chromatin, nucleosomes and chromatin have dual functions of structure and function. The position of nucleosomes on genomic DNA and the local structure of chromatin play a role in regulating a variety of biological processes including DNA replication, transcription, repair, recombination and splicing, and are mainly embodied in the following aspects:
(1) nucleosomes provide the most basic building blocks for the high in vivo compression of genomic DNA;
(2) dynamic change of nucleosome on genome DNA regulates and controls DNA replication, repair, recombination and other biological processes taking chromatin DNA as a template;
(3) the formation of nucleosomes affects the action of trans-acting factors with nucleosome DNA;
(4) the open or compressed state of chromatin local structures regulates the transcriptional activity of genes.
Therefore, the capability of assembling nucleosomes and chromatin structures at specific sites of a DNA sequence has very important influence on the expression regulation of genes, and the invention of the effective and convenient detection method has very important significance on the research on chromatin structure and the detection of chromatin related diseases.
Currently, there are several common methods for detecting nucleosome occupancy and local chromatin structure:
ChIP-seq method: the kit is suitable for large-scale detection in the whole genome range and the like, is expensive and has higher experimental operation requirements, and is not suitable for detecting the occupancy rate of nucleosomes at a few specific sites.
The Tilling-PCR method: a large amount of nested primers need to be designed, the experimental steps are complicated, and the price is high.
Sucrose gradient density centrifugation or analytical ultracentrifugation: open or compact structures suitable for detecting local chromatin structure, and not the nucleosome occupancy at a fixed point.
Disclosure of Invention
The present embodiments provide a method for detecting the ability to assemble chromatin structure at a specific site in a DNA sequence, so as to solve the above-mentioned problems.
The embodiment of the application adopts the following technical scheme:
a method for detecting the ability to assemble a chromatin structure at a specific site in a DNA sequence, comprising:
assembling a chromatin structure on a defined sample DNA sequence, the sample DNA sequence having a specific site thereon;
carrying out restriction enzyme digestion on the sampled DNA sequence assembled with the chromatin structure;
carrying out protease treatment on the sampled DNA sequence subjected to enzyme digestion treatment, and carrying out electrophoretic analysis;
and calculating the nucleosome occupancy rate of the specific site of the sampling DNA sequence according to the electrophoretic analysis result.
Optionally, the specific sites are: and the single enzyme cutting site is positioned in the range of 45bp upstream and downstream of the detection site and is provided with restriction enzyme.
Optionally, the chromatin structure is an in vitro assembled chromatin structure, comprising in particular: a circular chromatin structure and a linear chromatin structure.
Optionally, the restriction enzyme is: detecting the endonuclease corresponding to the locus.
Optionally, performing protease treatment on the sample DNA sequence after the digestion treatment, specifically including:
and (3) degrading the sampled DNA sequence subjected to enzyme digestion treatment by adopting protease through histone and endonuclease.
Optionally, the protease is specifically protease K, and 100-300 μ g of protease K is added into an experimental sample with the dosage of 1 μ g DNA, the temperature required by the reaction is 50-60 ℃, and the time required by the reaction is 30-60 min.
Optionally, a circular chromatin structure is assembled to the specific site of the sample DNA sequence;
calculating the nucleosome occupancy rate of the specific site of the sampled DNA sequence according to the electrophoretic analysis result, which specifically comprises the following steps:
two bands will form after electrophoresis, aboveThe band is the band which is digested by enzyme digestion, and the brightness is defined as A; the lower band is the band not digested by the enzyme, and the intensity is located as B. The occupancy rate of nucleosome of the detection site is
Optionally, a linear chromatin structure is assembled to the specific site of the sampled DNA sequence;
calculating the nucleosome occupancy rate of the specific site of the sampled DNA sequence according to the electrophoretic analysis result, which specifically comprises the following steps:
three bands can be formed after electrophoresis, the uppermost band is a band which is not digested by enzyme digestion, the sequence length is M1, and the brightness is A; the middle band is a fragment 1 digested by enzyme digestion, the sequence length is M2, and the brightness is B; the bottom band is fragment 2 after digestion, the sequence length is M3, and the brightness is C. The occupancy rate of nucleosomes at the detection sites is as follows:
the embodiment of the application adopts at least one technical scheme which can achieve the following beneficial effects:
in the technical scheme of the application, the specific site assembled with the DNA sequence with the chromatin structure is subjected to enzyme digestion, protease treatment and electrophoretic analysis only by adopting the conventional restriction endonuclease, the occupancy rate of nucleosomes (which can be understood as the chromatin structure) of the detection site (namely the specific site) can be calculated according to the electrophoretic analysis result, and then the efficiency of assembling the chromatin structure at the specific site on the DNA sequence and the assembling capacity of the nucleosomes are analyzed.
Drawings
The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this application, illustrate embodiment(s) of the application and together with the description serve to explain the application and not to limit the application. In the drawings:
FIG. 1 is a schematic diagram illustrating the steps of a method for detecting the ability to assemble a chromatin structure at a specific site in a DNA sequence according to an embodiment of the present disclosure;
FIG. 2 is a graph showing the results of examining the ability of specific sites on a circular DNA sequence to assemble chromatin structure;
FIG. 3 is a graph showing the results of examining the ability of specific sites on a linear DNA sequence to assemble chromatin structure.
Detailed Description
In order to make the objects, technical solutions and advantages of the present application more apparent, the technical solutions of the present application will be described in detail and completely with reference to the following specific embodiments of the present application and the accompanying drawings. It should be apparent that the described embodiments are only some of the embodiments of the present application, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present application.
It should be noted that the chromatin structure assembly ability detection scheme according to the present application can be applied to the detection of chromatin structure-related diseases, and can also be applied to the scientific research in the fields related to chromatin structure, epigenetics, structure biology, and for the fixed-point detection of the nucleosome occupancy rate and the local chromatin structure of a DNA sequence.
The technical solutions provided by the embodiments of the present application are described in detail below with reference to the accompanying drawings.
As shown in fig. 1, a schematic diagram of steps of a method for detecting the ability of assembling a chromatin structure at a specific site of a DNA sequence provided in the embodiments of the present application mainly includes the following steps:
step 11: assembling a chromatin structure on a defined sample DNA sequence, the sample DNA sequence having a specific site thereon;
in this step 11, a chromatin structure is assembled on the determined DNA sequence, which may be specifically realized by an assembly method such as a salt dialysis method or a chromatin-dependent remodeling factor method. Indeed, it is to be noted that reference to chromatin structure in the present application is also to be understood as reference to nucleosomes within the chromatin structure; the ability test performed in this application is not limited to the ability to assemble chromosomal structures, but also includes the ability to assemble nucleosomes.
Alternatively, in the present application, the specific site involved in the sampled DNA sequence is a single cleavage site with restriction enzymes located within 45bp upstream and downstream of the detection site.
Optionally, the chromatin structure is an in vitro assembled chromatin structure, comprising in particular: a circular chromatin structure and a linear chromatin structure.
Step 12: carrying out restriction enzyme digestion on the sampled DNA sequence assembled with the chromatin structure;
optionally, in this step, the restriction enzymes are: detecting the endonuclease corresponding to the locus.
Step 13: carrying out protease treatment on the sampled DNA sequence subjected to enzyme digestion treatment, and carrying out electrophoretic analysis;
specifically, in step 13, a protease may be used to perform histone degradation and endonuclease degradation on the sample DNA sequence after the digestion treatment, so as to achieve the purpose of removing protein. Considering that DNA sequences are protected from restriction endonuclease action after assembly of chromatin structure: that is, the site which is not assembled with the chromatin structure is cut after enzyme digestion, and the site which is assembled with the chromatin structure is not cut, and the electrophoresis detection analysis can be carried out subsequently.
When the protease treatment is carried out, the used protease is specifically protease K, 100-300 mu g of protease K is added into an experimental sample with the dosage of 1 mu g of DNA, the temperature required by the reaction is 50-60 ℃, and the time required by the reaction is 30-60 min.
Step 14: and calculating the nucleosome occupancy rate of the specific site of the sampling DNA sequence according to the electrophoretic analysis result.
Specifically, in calculating the nucleosome occupancy rate at a specific site, different treatments may be performed depending on the type of chromatin structure:
the first method is as follows: assembling a circular chromatin structure to the specific site of the sampled DNA sequence;
after electrophoresisTwo bands are formed, the upper band is the band which is digested by enzyme digestion, and the brightness is defined as A; the lower band is the band not digested by the enzyme, and the intensity is located as B. The occupancy rate of nucleosome of the detection site is
The second method comprises the following steps: assembling a linear chromatin structure to the specific site of the sampled DNA sequence;
three bands can be formed after electrophoresis, the uppermost band is a band which is not digested by enzyme digestion, the sequence length is M1, and the brightness is A; the middle band is a fragment 1 digested by enzyme digestion, the sequence length is M2, and the brightness is B; the bottom band is fragment 2 after digestion, the sequence length is M3, and the brightness is C. The occupancy rate of nucleosomes at the detection sites is as follows:
in fact, after the occupancy rate of nucleosomes at the detection site is calculated, the assembly capacity of local (specific site) chromatin structure and the like can be judged according to the occupancy rate, thereby realizing the capacity detection of assembling chromatin structure or assembling nucleosomes.
Through the technical scheme, the specific site of the DNA sequence assembled with the chromatin structure is subjected to enzyme digestion, protease treatment and electrophoretic analysis only by adopting the conventional restriction endonuclease, the occupancy rate of nucleosomes (which can be understood as the chromatin structure) of the detection site (namely the specific site) can be calculated according to the electrophoretic analysis result, and further, the efficiency of assembling the chromatin structure at the specific site on the DNA sequence and the assembly capacity of the nucleosomes are analyzed.
The detection scheme to which the present application relates is discussed in detail below by way of two specific examples.
Example 1: detection of the ability of a nucleosome to assemble at a specific site of reconstituted chromatin on a circular plasmid (1) the structure of reconstituted chromatin on a circular plasmid. The purified circular plasmid pUC601 was mixed with the purified histone octamer separately according to the following Table 1, and dialyzed against TE solution containing 2mol/L NaCl. In the dialysis process, TE is pumped into the dialysate to dilute the NaCl concentration in the dialysate to 0.6mol/L, and the dialysis process is controlled for 16 hours; the dialysis solution was replaced with TE buffer solution containing no NaCl and the dialysis was continued for 3 hours. Wherein, the reaction temperature of the whole experimental process is controlled to be 4 ℃.
TABLE 1
(2) And (4) carrying out restriction enzyme digestion. The occupancy of nucleosomes near the Nde I single cleavage site on the pUC601 plasmid sequence was examined. 1/5 of the dialyzed sample was added with 8U of Nde I and digested at 37 ℃ for 1 hour.
(3) Removing protein, and performing electrophoretic analysis. Adding 200 μ g protease K into the sample, digesting at 55 deg.C for 30min, adding phenol chloroform (25:24) of the same volume, and extracting; taking the supernatant, adding chloroform isoamyl alcohol (24:1) with the same volume for extraction; taking the supernatant, adding 1/10 volume of 3mol/L NaAc and 4 times volume of absolute ethyl alcohol, and standing at-80 ℃ for 30 min; centrifuging at 12000rpm for 10 min; removing supernatant, washing with 70% ethanol, and centrifuging at 12000rpm for 5 min; removing supernatant, and naturally drying; add 10. mu.l TE buffer to dissolve. 0.8% agarose gel electrophoresis, EB staining and photography.
(4) Calculating the nucleosome occupancy rate of the specific site and judging the local chromatin structure. Electrophoresis results A in FIG. 2 shows the chromatin structure assembled on the circular DNA plasmid detected by electron microscopy; b is restriction endonuclease cutting diagram; panel C shows the results of restriction endonuclease cleavage followed by electrophoresis, with lane 0 showing a control sample without assembled chromatin structure and lanes 1-3 showing assembled chromatin structure samples. Two bands appear in the sample in the lane, the upper band is the band after enzyme digestion, the nucleosome is not assembled at the site, and the brightness is defined as A; the lower band is the sample assembled with nucleosomes, which are not cut by endonucleases due to the protective effect of the nucleosomes. The nucleosome occupancy rates of the detection sites in samples 1, 2, and 3 were calculated as: 13.94%, 34.14%, 92.25%.
Example 2: detection of nucleosome assembling ability of reconstructed chromatin specific site on linear DNA sequence
(1) Chromatin structure was reconstructed on the linear DNA sequence. Plasmid pUC19 was digested with restriction enzyme Ssp I to prepare a linear DNA sequence. The cleaved and purified linearized plasmid pUC601 was mixed with the purified histone octamer, respectively, according to the following table, and dialyzed against TE solution containing 2mol/L NaCl. In the dialysis process, TE is pumped into the dialysate to dilute the NaCl concentration in the dialysate to 0.6mol/L, and the dialysis process is controlled for 16 hours; the dialysis solution was replaced with TE buffer solution containing no NaCl and the dialysis was continued for 3 hours. The whole experimental process is controlled to be carried out at 4 ℃.
TABLE 2
(2) And (4) carrying out restriction enzyme digestion. The occupancy of nucleosomes near the EcoR I single cleavage site on the pUC601 sequence was examined. 1/5 of the dialyzed sample was added with 12U of EcoR I and cleaved at 37 ℃ for 1 hour.
(3) Removing protein, and performing electrophoretic analysis. Adding 200 μ g protease K into the sample, digesting at 50 deg.C for 30min, adding phenol chloroform (25:24) of the same volume, and extracting; taking the supernatant, adding chloroform isoamyl alcohol (24:1) with the same volume for extraction; taking the supernatant, adding 1/10 volume of 3mol/L NaAc and 4 times volume of absolute ethyl alcohol, and standing at-80 ℃ for 30 min; centrifuging at 12000rpm for 10 min; removing supernatant, washing with 70% ethanol, and centrifuging at 12000rpm for 5 min; removing supernatant, and naturally drying; add 10. mu.l TE buffer to dissolve. Electrophoresis was performed on a 1% agarose gel and photographed by EB staining.
(4) Calculating the nucleosome occupancy rate of the specific site and judging the local chromatin structure. Electrophoresis results A in FIG. 3 shows the chromatin structure assembled on the linear DNA plasmid detected by electron microscopy; b is restriction endonuclease cutting diagram; panel C shows the results of restriction endonuclease cleavage followed by electrophoresis, with lane 0 showing a control sample without assembled chromatin structure and lanes 1-3 showing assembled chromatin structure samples. Three bands appear in the sample in the lane, the uppermost band is a band which is not digested by enzyme, the sequence length is M1, and the brightness is A; the middle band is a fragment 1 digested by enzyme digestion, the sequence length is M2, and the brightness is B; the bottom band is fragment 2 after digestion, the sequence length is M3, and the brightness is C. The occupancy rates of nucleosomes of the detection sites of the samples 1, 2 and 3 are respectively as follows: 57.34%, 81.56%, 84.38%.
The above description is only an example of the present application and is not intended to limit the present application. Various modifications and changes may occur to those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.
Claims (3)
1. A method for detecting the ability to assemble a chromatin structure at a specific site in a DNA sequence, comprising:
assembling a chromatin structure on a defined sample DNA sequence, the sample DNA sequence having a specific site thereon; the specific sites are: single restriction enzyme sites which are positioned in the range of 45bp upstream and downstream of the detection sites and are provided with restriction enzymes;
carrying out restriction enzyme digestion on the sampled DNA sequence assembled with the chromatin structure; the chromatin structure is an in vitro assembled chromatin structure, which specifically comprises: a circular chromatin structure and a linear chromatin structure; the restriction enzymes are: detecting endonuclease corresponding to the locus;
carrying out protease treatment on the sampled DNA sequence subjected to enzyme digestion treatment, and carrying out electrophoretic analysis; the protease is specifically protease K, 100-300 mu g of protease K is added into an experimental sample with the dosage of 1 mu g of DNA, the temperature required by the reaction is 50-60 ℃, and the time required by the reaction is 30-60 min;
and calculating the nucleosome occupancy rate of the specific site of the sampling DNA sequence according to the electrophoretic analysis result.
2. The detection method according to claim 1,
assembling a circular chromatin structure to the specific site of the sampled DNA sequence;
calculating the nucleosome occupancy rate of the specific site of the sampled DNA sequence according to the electrophoretic analysis result, which specifically comprises the following steps:
two bands can be formed after electrophoresis, the upper band is a band which is digested by enzyme digestion, and the brightness is defined as A; the lower band is the band not digested by the enzyme, the intensity is located as B, and the occupancy rate of nucleosomes at the detection site is
。
3. The detection method according to claim 1,
assembling a linear chromatin structure to the specific site of the sampled DNA sequence;
calculating the nucleosome occupancy rate of the specific site of the sampled DNA sequence according to the electrophoretic analysis result, which specifically comprises the following steps:
three bands can be formed after electrophoresis, the uppermost band is a band which is not digested by enzyme digestion, the sequence length is M1, and the brightness is A; the middle band is a fragment 1 digested by enzyme digestion, the sequence length is M2, and the brightness is B; the lowest band is the fragment 2 after enzyme digestion, the sequence length is M3, the brightness is C, and the occupancy rate of the nucleosome at the detection site is as follows:
。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710903858.2A CN107541558B (en) | 2017-09-29 | 2017-09-29 | Method for detecting capability of assembling chromatin structure on specific site of DNA sequence |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710903858.2A CN107541558B (en) | 2017-09-29 | 2017-09-29 | Method for detecting capability of assembling chromatin structure on specific site of DNA sequence |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107541558A CN107541558A (en) | 2018-01-05 |
| CN107541558B true CN107541558B (en) | 2021-03-23 |
Family
ID=60963914
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710903858.2A Active CN107541558B (en) | 2017-09-29 | 2017-09-29 | Method for detecting capability of assembling chromatin structure on specific site of DNA sequence |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107541558B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110938605B (en) * | 2019-11-20 | 2021-05-04 | 中山大学附属第五医院 | In vitro assembled HBV cccDNA nucleosome and preparation method thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2861255A2 (en) * | 2012-06-19 | 2015-04-22 | The Broad Institute, Inc. | Diagnostic and treatment methods in subjects having or at risk of developing resistance to cancer therapy |
| CN105695571A (en) * | 2016-01-29 | 2016-06-22 | 苏州金唯智生物科技有限公司 | DNA quantitative method based on rolling circle amplification |
| CN106566828A (en) * | 2016-11-11 | 2017-04-19 | 中国农业科学院农业基因组研究所 | Efficient whole-genome chromosome conformation capture technology (eHi-C) |
-
2017
- 2017-09-29 CN CN201710903858.2A patent/CN107541558B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2861255A2 (en) * | 2012-06-19 | 2015-04-22 | The Broad Institute, Inc. | Diagnostic and treatment methods in subjects having or at risk of developing resistance to cancer therapy |
| CN105695571A (en) * | 2016-01-29 | 2016-06-22 | 苏州金唯智生物科技有限公司 | DNA quantitative method based on rolling circle amplification |
| CN106566828A (en) * | 2016-11-11 | 2017-04-19 | 中国农业科学院农业基因组研究所 | Efficient whole-genome chromosome conformation capture technology (eHi-C) |
Non-Patent Citations (5)
| Title |
|---|
| DNAase footpHnnting: a simple method for the detection of protein-DNA binding specificity;David J.Gala等;《Nucleic Acids Research》;19780930;第5卷(第9期);第3157-3170页 * |
| High-Resolution Mapping and Characterization of Open Chromatin across the Genome;Alan P. Boyle等;《Cell》;20080125;第132卷;第311-322页 * |
| Measuring Arabidopsis Chromatin Accessibility Using DNase I-Polymerase Chain Reaction and DNase I-Chip Assays;Huan Shu等;《Breakthrough Technologies》;20130831;第162卷;第1794-1801页 * |
| 人类染色体隔离子区域核小体占据情况的统计分析;任松叶;《应用技术》;20120630(第221期);第1-3页 * |
| 依赖DNA序列的核小体定位的实验研究;柴荣;《中国优秀硕士学位论文全文数据库 基础科学辑》;20140515(第5期);第1-66页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107541558A (en) | 2018-01-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Unnikrishnan et al. | Dynamic changes in histone acetylation regulate origins of DNA replication | |
| Zhou et al. | CRISPR/Cas9-mediated efficient targeted mutagenesis of RAS in Salvia miltiorrhiza | |
| Frazee et al. | Ballgown bridges the gap between transcriptome assembly and expression analysis | |
| Schalamun et al. | Harnessing the MinION: An example of how to establish long‐read sequencing in a laboratory using challenging plant tissue from Eucalyptus pauciflora | |
| Staals et al. | RNA targeting by the type III-A CRISPR-Cas Csm complex of Thermus thermophilus | |
| Zhao et al. | A new look at adenovirus splicing | |
| Zeng et al. | Identification and analysis of ribosome-associated lncRNAs using ribosome profiling data | |
| McCartney‐Melstad et al. | Exon capture optimization in amphibians with large genomes | |
| Chalhoub et al. | Efficient cloning of plant genomes into bacterial artificial chromosome (BAC) libraries with larger and more uniform insert size | |
| WO2022052211A1 (en) | Homologous type 2 crispr/cas9 gene editing system and construction method therefor | |
| Chionh et al. | A multidimensional platform for the purification of non-coding RNA species | |
| Malousi et al. | DNA hypermethylation of alternatively spliced and repeat sequences in humans | |
| Soriano et al. | Different nucleosomal architectures at early and late replicating origins in Saccharomyces cerevisiae | |
| Xiong et al. | Simultaneous isolation of DNA, RNA, and protein from Medicago truncatula L. | |
| CN107025385A (en) | A kind of design method of circular rna primer | |
| CN107541558B (en) | Method for detecting capability of assembling chromatin structure on specific site of DNA sequence | |
| Steube et al. | High-intensity UV laser ChIP-seq for the study of protein-DNA interactions in living cells | |
| Chapple et al. | Characterization of the rat developmental liver transcriptome | |
| Stahl et al. | Simian virus 40 large tumor antigen on replicating viral chromatin: tight binding and localization on the viral genome | |
| Chen et al. | Symmetrical modification within a nucleosome is not required globally for histone lysine methylation | |
| Doyon et al. | Preparation and analysis of native chromatin-modifying complexes | |
| Zanini et al. | A bioinformatics pipeline for the analysis and target prediction of rna effectors in bidirectional communication during plant–microbe interactions | |
| Hollerer et al. | Strategies and Challenges in Identifying Function for Thousands of sORF‐Encoded Peptides in Meiosis | |
| Liang et al. | Protein methylome analysis in Arabidopsis reveals regulation in RNA-related processes | |
| Sahab et al. | Isoelectric point-based prefractionation of proteins from crude biological samples prior to two-dimensional gel electrophoresis |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |