CN104498594A - TALEs double-recognition detection method and application thereof - Google Patents
TALEs double-recognition detection method and application thereof Download PDFInfo
- Publication number
- CN104498594A CN104498594A CN201410726245.2A CN201410726245A CN104498594A CN 104498594 A CN104498594 A CN 104498594A CN 201410726245 A CN201410726245 A CN 201410726245A CN 104498594 A CN104498594 A CN 104498594A
- Authority
- CN
- China
- Prior art keywords
- tales
- albumen
- detection method
- double
- recognition detection
- 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.)
- Pending
Links
- 238000001514 detection method Methods 0.000 title claims abstract description 24
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 24
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 16
- 102000002260 Alkaline Phosphatase Human genes 0.000 claims abstract description 14
- 108020004774 Alkaline Phosphatase Proteins 0.000 claims abstract description 14
- 238000003745 diagnosis Methods 0.000 claims abstract description 6
- 239000000758 substrate Substances 0.000 claims abstract description 6
- 239000013613 expression plasmid Substances 0.000 claims abstract description 4
- 230000009465 prokaryotic expression Effects 0.000 claims description 10
- 241000894006 Bacteria Species 0.000 claims description 4
- 241000588724 Escherichia coli Species 0.000 claims description 4
- 238000001042 affinity chromatography Methods 0.000 claims description 4
- 238000010828 elution Methods 0.000 claims description 4
- 239000012149 elution buffer Substances 0.000 claims description 4
- 239000012145 high-salt buffer Substances 0.000 claims description 4
- 239000007788 liquid Substances 0.000 claims description 4
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 claims description 4
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 claims description 3
- 230000002934 lysing effect Effects 0.000 claims description 3
- 239000006228 supernatant Substances 0.000 claims description 3
- 238000005406 washing Methods 0.000 claims description 3
- 108020004414 DNA Proteins 0.000 abstract description 14
- 238000000034 method Methods 0.000 abstract description 6
- 108091028043 Nucleic acid sequence Proteins 0.000 abstract description 5
- 238000011160 research Methods 0.000 abstract description 5
- 230000035945 sensitivity Effects 0.000 abstract description 4
- 238000003759 clinical diagnosis Methods 0.000 abstract description 3
- 206010064571 Gene mutation Diseases 0.000 abstract 1
- 238000012408 PCR amplification Methods 0.000 abstract 1
- 239000011248 coating agent Substances 0.000 abstract 1
- 238000000576 coating method Methods 0.000 abstract 1
- 239000000523 sample Substances 0.000 description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 6
- 150000002460 imidazoles Chemical class 0.000 description 6
- 230000008878 coupling Effects 0.000 description 4
- 238000010168 coupling process Methods 0.000 description 4
- 238000005859 coupling reaction Methods 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 230000004568 DNA-binding Effects 0.000 description 3
- 235000011187 glycerol Nutrition 0.000 description 3
- 239000011780 sodium chloride Substances 0.000 description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- 108010077850 Nuclear Localization Signals Proteins 0.000 description 2
- 108010073062 Transcription Activator-Like Effectors Proteins 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 108020004707 nucleic acids Proteins 0.000 description 2
- 102000039446 nucleic acids Human genes 0.000 description 2
- 150000007523 nucleic acids Chemical class 0.000 description 2
- 230000029279 positive regulation of transcription, DNA-dependent Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- 108090001008 Avidin Proteins 0.000 description 1
- 101150039808 Egfr gene Proteins 0.000 description 1
- 241000672609 Escherichia coli BL21 Species 0.000 description 1
- 101500025419 Homo sapiens Epidermal growth factor Proteins 0.000 description 1
- 208000009869 Neu-Laxova syndrome Diseases 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-N ammonia Natural products N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000002038 chemiluminescence detection Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 108700021358 erbB-1 Genes Proteins 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 229940116978 human epidermal growth factor Drugs 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 235000019799 monosodium phosphate Nutrition 0.000 description 1
- 230000000869 mutational effect Effects 0.000 description 1
- GVUGOAYIVIDWIO-UFWWTJHBSA-N nepidermin Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](NC(=O)[C@H](CS)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CS)NC(=O)[C@H](C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C(C)C)C(C)C)C1=CC=C(O)C=C1 GVUGOAYIVIDWIO-UFWWTJHBSA-N 0.000 description 1
- 230000009871 nonspecific binding Effects 0.000 description 1
- 239000002773 nucleotide Substances 0.000 description 1
- 125000003729 nucleotide group Chemical group 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 1
- 229910000162 sodium phosphate Inorganic materials 0.000 description 1
- QTENRWWVYAAPBI-YCRXJPFRSA-N streptomycin sulfate Chemical compound OS(O)(=O)=O.OS(O)(=O)=O.OS(O)(=O)=O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O.CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](N=C(N)N)[C@H](O)[C@@H](N=C(N)N)[C@H](O)[C@H]1O QTENRWWVYAAPBI-YCRXJPFRSA-N 0.000 description 1
- 230000005945 translocation Effects 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a TALEs double-recognition detection method. The TALEs double-recognition detection method comprises the following steps: step 1, constructing TALEs prokaryotic protein expression plasmids; step 2, prokaryotically expressing TALEs proteins; step 3, coating the TALEs proteins on a microporous plate, adding a DAN sample, and capturing DNAs by the TALEs proteins coated on the microporous plate; step 4, recognizing the captured DNAs by the TALEs proteins marked with alkaline phosphatase, adding the substrate of alkaline phosphatase, generating a chemiluminescence signal, and interpreting the signal. According to the TALEs double-recognition detection method disclosed by the invention, the characteristic of the TALEs proteins capable of specifically recognizing the DNA sequence is utilized, and then the signal is amplified through chemiluminescence, thus the TALEs double-recognition detection method is high in sensitivity and capable of being used for rapid diagnosis. The TALEs double-recognition detection method disclosed by the invention is greatly different from the traditional detection means, does not have a middle PCR amplification step, and has a high detection sensitivity and a low false positive rate. The method can be used for both gene mutation detection and the like in scientific researches, and can also be used for clinical diagnosis.
Description
Technical field
The invention belongs to biology field, be specifically related to a kind of two recognition detection method of TALEs and application thereof.
Background technology
TALE (Transcription Activator Like Effectors) is a class DBP.Its DNA binding domains is made up of the repeating unit of variable number, natural TALEs is primarily of being positioned at the encoding transport signals (translocation signal) of N end, being positioned at nuclear localization signal (the nuclear localization signals of C end, and transcriptional activation domain (transcriptional activation domain, AD) and be positioned at middle DNA binding domains three part composition NLS).DNA binding domains is generally 34 by comprising 33 ~ 35() repeating unit of individual amino-acid residue is in series, mediated dna specific recognition and combination.Each repeating unit sequence high conservative, the key distinction be the 12nd and the 13rd repeat variable pair of residue (repeat variant di-residue, RVD).Research finds that the RVD of each repeating unit determines its Nucleotide identified, and a simple rule is followed in its identification: NI=A, HD=C, NG=T, NN=G or A.TALEs can identify any DNA sequence in theory.
Also be not used for the diagnosis of gene at present with TALEs albumen, the two recognition detection technology of TALEs of the present invention, does abrupt climatic change with it.The method not only may be used for the detection of scientific research, also may be used for clinical diagnosis.
Summary of the invention
the technical problem solved:
The object of the invention is to overcome the deficiencies in the prior art and the two recognition detection method of a kind of TALEs of developing, the method can detect different genes, point mutation etc., can be used for the research of molecular diagnosis, genetic expression, detection sensitivity of the present invention is higher, and false positive rate is lower.
technical scheme:
The present invention utilizes TALEs to identify the characteristic of DNA, expresses the TALEs albumen of specific recognition DNA sequence dna, then identifying that the albumen of different DNA sequence dna is coated on microwell plate.Subsequently add DNA sample and the TALEs protein binding be coated on microwell plate, then add the TALEs albumen that coupling has alkaline phosphatase again, this albumen specific recognition DNA sequence dna, finally adds the substrate of alkaline phosphatase, chemiluminescence detection.Thus detect different genes, point mutation etc.
The two recognition detection method of TALEs, step is as follows:
Step one: the structure of TALEs prokaryotic protein expression plasmid: cut pET-16b carrier with NdeI and BamHI, then NdeI and the BamH site of TALEs sequence clone to pET-16b carrier, obtains pET-16b-TALEs prokaryotic expression carrier;
Step 2: prokaryotic expression TALEs albumen: pET-16b-TALEs prokaryotic expression carrier step one prepared is transformed in e. coli bl21, IPTG induces, collect bacterium and lysing cell, centrifuging and taking supernatant liquor, carries out Ni post affinity chromatography, after treating that target protein is combined with Ni post, with rinsing liquid washing, rear elution buffer rinses, and the high-salt buffer in elution fraction is replaced as PBS solution, finally carry out the detection of SDS-PAGE, gained is the TALEs albumen of purifying.
Step 3: the directed high-density of TALEs albumen is coated on microwell plate.Add the DNA sample of extraction, DNA is coated on the TALEs albumen Packet capturing on microwell plate.
Step 4: the DNA having the TALEs albumen identification of alkaline phosphatase captured with coupling, adds the substrate of alkaline phosphatase, produce chemiluminescence signal, and interpretation signal.
The two application of recognition detection method in gene diagnosis of described TALEs.
beneficial effect
The present invention breaks the nucleic acid bind nucleic acid of gene chip, the characteristic of albumen identification albumen in protein chip, with protein-specific identification DNA, stable, and remolding sensitivity is higher, may be used for quick diagnosis.Compared with prior art, advantage of the present invention is can the different gene of high throughput testing and multiple mutational site, and the method not only may be used for the detection of scientific research, also can be used as clinical diagnosis.
Accompanying drawing explanation
Fig. 1 is the signal results figure detected with chemoluminescence method, and as shown in Figure 1, TALEs detects the detected result in EGFR gene T790M site to result.
Embodiment
By reference to the accompanying drawings the present invention is described in further detail below by embodiment.But it will be understood to those of skill in the art that the following example only for illustration of the present invention, and should not be considered as limiting scope of the present invention.Unreceipted concrete technology or condition person in embodiment, according to the technology described by the document in this area or condition or carry out according to product description.Agents useful for same or the unreceipted production firm person of instrument, being can by the conventional products of commercial acquisition.
embodiment 1
1, material: pET-16b is purchased from Novagen, e. coli bl21 is purchased from Takara, and the TALEs albumen being marked with alkaline phosphatase is marked by Beijing Bo Aosen Bioisystech Co., Ltd.
2, method
Step one: the structure of TALEs prokaryotic protein expression plasmid: cut PET-16b carrier with NdeI and BamHI, then NdeI and the BamH site of TALEs sequence clone to pET-16b carrier, obtains pET-16b-TALEs prokaryotic expression carrier;
Step 2: prokaryotic expression TALEs albumen: pET-16b-TALEs prokaryotic expression carrier step one prepared is transformed into e. coli bl21, IPTG induces, collect bacterium and lysing cell (50mM NaH2PO4, pH 8.0,300mM NaCl, 10mM imidazoles), centrifuging and taking supernatant liquor, carry out Ni post affinity chromatography, after treating that target protein is combined with Ni post, first use rinsing liquid (50mM NaH
2p0
4, pH 8.0,300mM NaCI, 20mM imidazoles) and washing, use elution buffer (50mM NaH afterwards
2p0
4, pH8.0,300mM NaCI, 250mM imidazoles) rinse, by high-salt buffer (the 50mM NaH in elution fraction
2p0
4, pH8.0,300mM NaCI, 250mM imidazoles) and be replaced as PBS solution, finally carry out the detection of SDS-PAGE, gained is the TALEs albumen of purifying;
Step 3: TALEs albumen orientation is coated on microwell plate, adds DNA sample, DNA is coated on the TALEs albumen Packet capturing on microwell plate;
Step 4: with the DNA that the TALEs albumen identification being marked with alkaline phosphatase is captured, add the substrate of alkaline phosphatase, produce chemiluminescence signal, and interpretation signal.
embodiment 2
Detect the sudden change of the T790M of human epidermal growth factor acceptor (EGFR) gene 20 exon
1. the TALEs probe of selective recognition wild-type T790 sequence: TALEs-T790(CGACATCACGCAGCTC) (SEQ ID NO.1), identify the TALEs probe of the sequence of 790M: TALEs-790M(CGACATCATGCAGCTC) (SEQ ID NO.2), identify the TALEs probe of negative control sequence: TALEs-control(CGACATCTCGTAGCTC) (SEQ ID NO.3) build the recombinant plasmid pET-16b-TALEs-T790 (CGACATCACGCAGCTC) of prokaryotic expression, pET-16b-TALEs-790M E (CGACATCATGCAGCTC) and pET-16b-TALEs-control(CGACATCTCGTAGCTC).These three vector intestinal bacteria E. coli BL21: transformed bacteria overnight incubation in the substratum of 5ml ammonia benzyl resistance, be transferred to next day in the identical substratum of 500ml and cultivate, through IPTG16 DEG C of induction of spending the night, collect the TALEs-790M obtained, TALEs-790M and TALEs-control albumen thalline, the albumen of centrifugal acquisition solubility after high pressure fragmentation, carries out Ni post affinity chromatography.After treating that target protein is combined with Ni post, first use rinsing liquid (20mM Tris-HCl, 500mM NaCl, 5%(v/v) glycerine, 60mM imidazoles, pH8.0) wash, use elution buffer (20mM Tris-HCl, 500mM NaCl, 5%(v/v) glycerine afterwards, 500mM imidazoles, pH8.0) wash away albumen.PD-10 desalting column is utilized to be replaced as the high-salt buffer in elution fraction containing 20%(volume percent) PBS solution of glycerine, carry out the detection of SDS-PAGE afterwards, gained is the TALEs-T790 of purifying, TALEs-790M and TALEs-control albumen.
2.TALEs-T790 is coated on microwell plate.With being marked with the TALEs-T790 albumen of avidin and combining with 9 orifice plates of Streptomycin sulphate, TALEs-T790 albumen is just coated on microwell plate.
3. add the DNA sample extracted and carry out hybrid capture, remove the DNA of non-specific binding.Then the TALEs-790M albumen of alkaline phosphatase there is is to join IC1 coupling, in IC2 hole, there is the TALEs-control albumen of alkaline phosphatase to join in other holes coupling, finally add the substrate of alkaline phosphatase, produce chemiluminescence signal, and interpretation signal.
Table 1 sequence
| Sequence | SEQ ID NO. | |
| Identify the TALEs-T790 probe of wild-type T790 sequence | CGACATCACGCAGCTC | 1 |
| Identify the TALEs-790M probe of the sequence of 790M | CGACATCATGCAGCTC | 2 |
| Identify that the TALEs-control of negative control sequence visits | CGACATCTCGTAGCTC | 3 |
Claims (2)
1.TALEs two recognition detection method, it is characterized in that, step is as follows:
Step one: the structure of TALEs prokaryotic protein expression plasmid: cut pET-16b carrier with NdeI and BamHI, then NdeI and the BamH site of TALEs sequence clone to pET-16b carrier, obtains pET-16b-TALEs prokaryotic expression carrier;
Step 2: prokaryotic expression TALEs albumen: pET-16b-TALEs prokaryotic expression carrier step one prepared is transformed in e. coli bl21, IPTG induces, collect bacterium and lysing cell, centrifuging and taking supernatant liquor, carries out Ni post affinity chromatography, after treating that target protein is combined with Ni post, with rinsing liquid washing, rear elution buffer rinses, and the high-salt buffer in elution fraction is replaced as PBS solution, finally carry out the detection of SDS-PAGE, gained is the TALEs albumen of purifying;
Step 3: TALEs albumen orientation is coated on microwell plate, adds DNA sample, DNA is coated on the TALEs albumen Packet capturing on microwell plate;
Step 4: with the DNA that the TALEs albumen identification being marked with alkaline phosphatase is captured, add the substrate of alkaline phosphatase, produce chemiluminescence signal, and interpretation signal.
2. the two application of recognition detection method in gene diagnosis of TALEs according to claim 1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410726245.2A CN104498594A (en) | 2014-12-04 | 2014-12-04 | TALEs double-recognition detection method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410726245.2A CN104498594A (en) | 2014-12-04 | 2014-12-04 | TALEs double-recognition detection method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104498594A true CN104498594A (en) | 2015-04-08 |
Family
ID=52940044
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410726245.2A Pending CN104498594A (en) | 2014-12-04 | 2014-12-04 | TALEs double-recognition detection method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104498594A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108949914A (en) * | 2018-07-06 | 2018-12-07 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Escherichia coli O 157 based on activating transcription factor sample effector nuclease: H7 detection kit and detection method |
| CN110079583A (en) * | 2019-05-27 | 2019-08-02 | 重庆博艾迈迪森生物科技有限公司 | A kind of immunochromatography detection method, detection kit and its application of nucleic acid |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090068164A1 (en) * | 2005-05-05 | 2009-03-12 | The Ariz Bd Of Regents On Behald Of The Univ Of Az | Sequence enabled reassembly (seer) - a novel method for visualizing specific dna sequences |
| WO2013102290A1 (en) * | 2012-01-04 | 2013-07-11 | 清华大学 | Method for specifically recognizing dna containing 5-methylated cytosine |
| CN103233004A (en) * | 2013-04-28 | 2013-08-07 | 新疆农垦科学院 | Artificial DNA (deoxyribonucleic acid) molecule and method for detecting expression of target gene |
-
2014
- 2014-12-04 CN CN201410726245.2A patent/CN104498594A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20090068164A1 (en) * | 2005-05-05 | 2009-03-12 | The Ariz Bd Of Regents On Behald Of The Univ Of Az | Sequence enabled reassembly (seer) - a novel method for visualizing specific dna sequences |
| WO2013102290A1 (en) * | 2012-01-04 | 2013-07-11 | 清华大学 | Method for specifically recognizing dna containing 5-methylated cytosine |
| CN103233004A (en) * | 2013-04-28 | 2013-08-07 | 新疆农垦科学院 | Artificial DNA (deoxyribonucleic acid) molecule and method for detecting expression of target gene |
Non-Patent Citations (4)
| Title |
|---|
| 左伋: "《遗传医学进展》", 31 May 2014, 复旦大学出版社 * |
| 曹孟德等: "《HLA分子生物学及临床应用》", 31 December 1998, 河南医科大学出版社 * |
| 武峰: "《安全血液与输血检验技术》", 31 July 2007, 中国科学技术出版社 * |
| 马文丽: "《医学分子生物学》", 31 December 2013, 北京大学医学出版社 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108949914A (en) * | 2018-07-06 | 2018-12-07 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Escherichia coli O 157 based on activating transcription factor sample effector nuclease: H7 detection kit and detection method |
| CN108949914B (en) * | 2018-07-06 | 2021-11-30 | 军事科学院军事医学研究院环境医学与作业医学研究所 | Escherichia coli O157H 7 detection kit based on transcription activator-like effector nuclease and detection method |
| CN110079583A (en) * | 2019-05-27 | 2019-08-02 | 重庆博艾迈迪森生物科技有限公司 | A kind of immunochromatography detection method, detection kit and its application of nucleic acid |
| CN110079583B (en) * | 2019-05-27 | 2023-10-27 | 重庆博艾迈迪森生物科技有限公司 | Immunochromatography detection method and detection kit for nucleic acid and application of detection kit |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Pękowska et al. | Gain of CTCF-anchored chromatin loops marks the exit from naive pluripotency | |
| Chen et al. | Sensing self and foreign circular RNAs by intron identity | |
| US20220239615A1 (en) | Rna targeting methods and compositions | |
| Ohle et al. | Transient RNA-DNA hybrids are required for efficient double-strand break repair | |
| Ablasser et al. | Cell intrinsic immunity spreads to bystander cells via the intercellular transfer of cGAMP | |
| Chacko et al. | The lncRNA RZE1 controls cryptococcal morphological transition | |
| Cho et al. | Interchromosomal homology searches drive directional ALT telomere movement and synapsis | |
| JP2021000086A (en) | Methods and reagents for molecular proximity detection using rna-guided nucleic acid-binding proteins | |
| CN104651401B (en) | A kind of method that mir-505 diallele knocks out | |
| AU2014333776A1 (en) | Methods and kits for detecting nucleic acid sequences of interest using DNA-binding protein domain | |
| CN104131105A (en) | Method for screening aptamer specifically bound with alpha-fetoprotein | |
| CN109477132A (en) | Ribonucleic acid (RNA) interaction | |
| Mehta et al. | Temporal resolution of gene derepression and proteome changes upon PROTAC-mediated degradation of BCL11A protein in erythroid cells | |
| CN104498594A (en) | TALEs double-recognition detection method and application thereof | |
| CN115210370A (en) | RNA detection and transcription-dependent editing using reprogrammed tracrRNA | |
| CN109207518A (en) | Drug induced CRISPR/Cas9 system for gene transcriptional activation | |
| CN110004171A (en) | A kind of yeast two-hybrid method | |
| CN102766633A (en) | DNA (deoxyribonucleic acid) aptamer available for detecting HCC (hepatocellular carcinoma) cell line Bel-7404 and screening method and application thereof | |
| Arnould et al. | Loop extrusion as a mechanism for DNA Double-Strand Breaks repair foci formation | |
| US11345959B2 (en) | Method for exploring useful genetic resources through bulk metagenome analysis and use thereof | |
| CN108929918B (en) | On-site rapid detection method and kit for detecting PRRSV (porcine reproductive and respiratory syndrome Virus) | |
| Miranda et al. | Deciphering interactions used by the influenza virus NS1 protein to silence the host antiviral sensor protein RIG-I using a bacterial reverse two-hybrid system | |
| CN104531633A (en) | Cas9-scForkI fusion protein and application thereof | |
| CN103993097B (en) | A kind of electrochemical detection method of micro ribonucleic acid | |
| Luo et al. | Yeast actin‐related protein Arp6 negatively regulates Agrobacterium‐mediated transformation of yeast cell |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C41 | Transfer of patent application or patent right or utility model | ||
| CB03 | Change of inventor or designer information |
Inventor after: Cheng Gang Inventor before: Li Yunying |
|
| COR | Change of bibliographic data | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20160713 Address after: 215000 Wuzhong District City, Suzhou Province, Guo Xiang street, East Ring Road, No. 2, building 999, No. Applicant after: Suzhou Kechuang Biotechnology Co.,Ltd. Address before: 510000, No. 45, Zhongshan Avenue, Tianhe District, Guangdong, Guangzhou, T41001 Applicant before: Li Yunying |
|
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20150408 |