CN102912022A - Method for detecting polymorphism of BCL2L11 gene - Google Patents
Method for detecting polymorphism of BCL2L11 gene Download PDFInfo
- Publication number
- CN102912022A CN102912022A CN2012104081189A CN201210408118A CN102912022A CN 102912022 A CN102912022 A CN 102912022A CN 2012104081189 A CN2012104081189 A CN 2012104081189A CN 201210408118 A CN201210408118 A CN 201210408118A CN 102912022 A CN102912022 A CN 102912022A
- Authority
- CN
- China
- Prior art keywords
- fragment
- primer
- primers
- amplification
- amplify
- 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.)
- Granted
Links
- 238000000034 method Methods 0.000 title claims abstract description 19
- 108010040168 Bcl-2-Like Protein 11 Proteins 0.000 title claims abstract description 14
- 239000012634 fragment Substances 0.000 claims abstract description 39
- 230000003321 amplification Effects 0.000 claims abstract description 19
- 238000003199 nucleic acid amplification method Methods 0.000 claims abstract description 19
- 238000006243 chemical reaction Methods 0.000 claims abstract description 18
- 238000012217 deletion Methods 0.000 claims abstract description 8
- 230000037430 deletion Effects 0.000 claims abstract description 8
- 238000001514 detection method Methods 0.000 claims description 12
- 238000001962 electrophoresis Methods 0.000 claims description 8
- 230000008034 disappearance Effects 0.000 claims description 7
- 238000002156 mixing Methods 0.000 claims description 6
- 238000007400 DNA extraction Methods 0.000 claims description 3
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 3
- 229920002684 Sepharose Polymers 0.000 claims description 3
- 239000003146 anticoagulant agent Substances 0.000 claims description 3
- 229940127219 anticoagulant drug Drugs 0.000 claims description 3
- 230000010100 anticoagulation Effects 0.000 claims description 3
- 230000004087 circulation Effects 0.000 claims description 3
- 238000002474 experimental method Methods 0.000 claims description 3
- 238000003384 imaging method Methods 0.000 claims description 3
- 239000012160 loading buffer Substances 0.000 claims description 3
- 210000005259 peripheral blood Anatomy 0.000 claims description 3
- 239000011886 peripheral blood Substances 0.000 claims description 3
- 238000004321 preservation Methods 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 238000012360 testing method Methods 0.000 claims description 3
- 108090000623 proteins and genes Proteins 0.000 abstract description 9
- 238000005516 engineering process Methods 0.000 abstract description 6
- 102000001765 Bcl-2-Like Protein 11 Human genes 0.000 description 10
- 108020004414 DNA Proteins 0.000 description 7
- 229940121358 tyrosine kinase inhibitor Drugs 0.000 description 6
- 239000005483 tyrosine kinase inhibitor Substances 0.000 description 6
- 150000004917 tyrosine kinase inhibitor derivatives Chemical class 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 101150092671 BIM gene Proteins 0.000 description 4
- 108091028043 Nucleic acid sequence Proteins 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 102000052116 epidermal growth factor receptor activity proteins Human genes 0.000 description 3
- 108700015053 epidermal growth factor receptor activity proteins Proteins 0.000 description 3
- YOHYSYJDKVYCJI-UHFFFAOYSA-N n-[3-[[6-[3-(trifluoromethyl)anilino]pyrimidin-4-yl]amino]phenyl]cyclopropanecarboxamide Chemical compound FC(F)(F)C1=CC=CC(NC=2N=CN=C(NC=3C=C(NC(=O)C4CC4)C=CC=3)C=2)=C1 YOHYSYJDKVYCJI-UHFFFAOYSA-N 0.000 description 3
- 208000032791 BCR-ABL1 positive chronic myelogenous leukemia Diseases 0.000 description 2
- 208000010833 Chronic myeloid leukaemia Diseases 0.000 description 2
- 102000001301 EGF receptor Human genes 0.000 description 2
- 108060006698 EGF receptor Proteins 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 208000033761 Myelogenous Chronic BCR-ABL Positive Leukemia Diseases 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- 239000003153 chemical reaction reagent Substances 0.000 description 2
- 230000004927 fusion Effects 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 208000002154 non-small cell lung carcinoma Diseases 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 208000029729 tumor suppressor gene on chromosome 11 Diseases 0.000 description 2
- WEVYNIUIFUYDGI-UHFFFAOYSA-N 3-[6-[4-(trifluoromethoxy)anilino]-4-pyrimidinyl]benzamide Chemical compound NC(=O)C1=CC=CC(C=2N=CN=C(NC=3C=CC(OC(F)(F)F)=CC=3)C=2)=C1 WEVYNIUIFUYDGI-UHFFFAOYSA-N 0.000 description 1
- 208000010839 B-cell chronic lymphocytic leukemia Diseases 0.000 description 1
- 208000003950 B-cell lymphoma Diseases 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000258955 Echinodermata Species 0.000 description 1
- 108700024394 Exon Proteins 0.000 description 1
- 102100030708 GTPase KRas Human genes 0.000 description 1
- 101000584612 Homo sapiens GTPase KRas Proteins 0.000 description 1
- 208000031422 Lymphocytic Chronic B-Cell Leukemia Diseases 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- 102000029749 Microtubule Human genes 0.000 description 1
- 108091022875 Microtubule Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 239000012807 PCR reagent Substances 0.000 description 1
- 102000001253 Protein Kinase Human genes 0.000 description 1
- 102000052575 Proto-Oncogene Human genes 0.000 description 1
- 108700020978 Proto-Oncogene Proteins 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- 241000713810 Rat sarcoma virus Species 0.000 description 1
- 108700025690 abl Genes Proteins 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 208000032852 chronic lymphocytic leukemia Diseases 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 210000004688 microtubule Anatomy 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 108060006633 protein kinase Proteins 0.000 description 1
- 102000027426 receptor tyrosine kinases Human genes 0.000 description 1
- 108091008598 receptor tyrosine kinases Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000008685 targeting Effects 0.000 description 1
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention provides a method for detecting polymorphism of a BCL2L11 gene. The method is characterized by comprising steps as follows: two pairs of primers are designed, namely a mutant type primer and a wild type primer; one pair of primers crosses a 2903bp deletion region, when a fragment is not deleted, a reaction is limited based on an experimental condition, so that a long fragment cannot be amplified; when the fragment is deleted, one 261bp fragment can be amplified; and another pair of primers crosses the 2903bp deletion region, when the fragment is not deleted, and one 365bp fragment can be amplified. According to the technology, two different products can be amplified in one same reaction system, so that a type of the gene of a sample can be identified; the amplified fragments are moderate in length, and an amplification condition is wide in range, and a used instrument is simple.
Description
Technical field:
The present invention relates to biotechnology and medical field, molecular biology especially, polymerase chain reaction.
Background technology:
Tyrosine kinase inhibitor (TKI) is an important class anticancer targeting medicine, in the unusual oncotherapy of kinases, be widely applied, and effect is very good, such as the chronic myelocytic leukemia (CML) of breaking point bunch Ji Qu-cell abl oncogene (BCR-ABL) fusion, the nonsmall-cell lung cancer (NSCLC) of EGF-R ELISA (Epidermal growth factor receptor, EGFR) exons 1 9/21 sudden change.But along with the crowd's of use continuous increase, find to have the patient treatment effect of portion gene sudden change not remarkable, effect is fine when perhaps having some patients were to begin, but along with treatment is carried out, the effect of medicine weakens gradually.Wherein the T790M of EGFR extron 20 sudden change is the important reason that causes drug resistance, continuous expansion along with research, find EGF-R ELISA (Epidermal growth factor receptor, EGFR) other sudden changes of path also can cause drug effect, such as kirsten rat sarcoma virus proto-oncogene (Kirsten rat sarcoma viral oncogene homolog, kras) codon 12,13,61 sudden changes, echinoderms microtubule sample albumen 4-anaplastic lymphoma receptor tyrosine kinase (EML4-ALK) fusion etc.Certainly, it below all is the special sudden change of cancer cells, and B-Cell Chronic Lymphocytic Leukemia/lymphoma-2 sample 11(BCL2L11) also the tolerance with the TKI medicine is relevant for gene (having another name called BIM) polymorphism, and it belongs to germline mutation, just can carry out by the atraumatic detection, can judge to the suitability of TKI medicine in early days that treatment cost is saved in more effective treatment.
The BIM gene is apoptosis-related genes, participates in the apoptotic process that TKI induces.When being suppressed of BIM, the effect of meeting remarkably influenced TKI when expression level reduces.The polymorphism of BIM refer in its gene intron 2 can disappearance 2903bp dna sequence dna, cause the splicing mistake of BIM exon 3 and 4, obtain incomplete protein, make the BIM can not functionating.Incomplete statistics, this polymorphism has reached 12.3% in the crowd of East Asia, but does not also find this kind variation in Africa and European crowd.
About the detection of BIM gene pleiomorphism, relevant patent does not also have at present, and bibliographical information is also very limited.The detection method that adopts in the existing report is the dna sequence dna that needs one section 4226bp of amplification, and still, long sequence like this is in actually operating and be not easy realization, and failed probability is very high, can not be developed as the stable detection means of BIM gene pleiomorphism.And in the time can only obtaining person under inspection's paraffin-embedded tissue, its DNA has been fractured into the following small segment of 500bp owing to living through the processing of chemical reagent, can't adopt this kind method to detect.
In view of above problem, a kind of invention of method for quick new, that operating performance is higher is imperative.
Summary of the invention:
The technical problem to be solved in the present invention mainly is:
The polymorphism of BIM refer in its gene intron 2 can disappearance 2903bp dna sequence dna, cause the splicing mistake of BIM exon 3 and 4, obtain incomplete protein, make the BIM can not functionating.
The detection method that adopts in the existing report is the dna sequence dna that needs one section 4226bp of amplification, and still, long sequence like this is in actually operating and be not easy realization, and failed probability is very high, can not be developed as the stable detection means of BIM gene pleiomorphism.
And in the time can only obtaining person under inspection's paraffin-embedded tissue, its DNA has been fractured into the following small segment of 500bp owing to living through the processing of chemical reagent, can't adopt this kind method to detect.
In order to overcome the above problems, the technology of the present invention provides a kind of method of the BCL2L11 of detection gene pleiomorphism, and it comprises following performing step:
Designed two pairs of primers, mutant primer and wild-type primer, pair of primers are crossed over the disappearance zone of 2903bp, when this fragment does not lack, and experiment condition limited reactions, the fragment of the length like this that can not increase; When this fragment deletion, can amplify the fragment of a 261bp;
Pair of primers when this fragment does not lack, can amplify the fragment of a 365bp in the disappearance zone of 2903bp in addition.
The wild-type primer sequence of described amplification is:
Forward primer wmF:CAGAACAGACACTGGAACAAAATGA
Reverse primer wR:GGGAGCATTTTCATGAGTACAACAA
The mutant primer sequence of described amplification is:
Forward primer wmF:CAGAACAGACACTGGAACAAAATGA(is identical with wild-type F)
Reverse primer mR:GGTGGCCATACAAATCTAAGCCAG
Described concrete treatment step is as follows:
Sample process is got peripheral blood 2ml in the EDTA anticoagulant tube, gently puts upside down mixing, 4 ℃ of preservations;
DNA extraction is got 200 μ L anticoagulations and is extracted DNA with test kit, adopts the running gel imaging that DNA purity and concentration are detected;
The configuration of reaction system is in the certain reaction system of the aerial adding of each reaction;
Increase by certain program;
The electrophoresis detection amplification.
Below the described reaction system:
Describedly increase by following program:
95℃3min
94 ℃ of 30sec → (50-60 ℃) all right 30sec → 72 ℃ 30sec, 40 circulations
72℃5min→4℃
Described electrophoresis detection amplification step is as follows: join 2% sepharose, get 9 μ L(3) loading behind the product of amplification and the 10 times of concentration sample-loading buffer mixings, 120V electrophoresis 30min, ultraviolet judgement PCR result; When fragment deletion, can amplify the fragment of a 261bp; When this fragment does not lack, can amplify the fragment of a 365bp; If 261bp and 365bp product exist simultaneously, this sample is heterozygote.
The invention has the beneficial effects as follows: this technology two different products that in same reaction system, increase simultaneously, identify the sample genotype.And expanding fragment length is moderate, and the amplification condition wide scope uses instrument simple.
Description of drawings:
Fig. 1 is the schematic diagram of three primer positions of the present invention;
Embodiment:
The present invention is applied to biotechnology and medical field, molecular biology especially, molecular diagnosis, Real-time quantitative PCR.
As shown in Figure 1, the present invention has designed two pairs of primers in order to overcome the above problems, and pair of primers is crossed over the disappearance zone of this 2903bp, when this fragment does not lack, and experiment condition limited reactions, the fragment of the length like this that can not increase; When this fragment deletion, can amplify the fragment of a 261bp.
Pair of primers when this fragment does not lack, can amplify the fragment of a 365bp in the zone of this 2903bp in addition;
Amplification wild-type primer sequence:
Forward primer wmF:CAGAACAGACACTGGAACAAAATGA
Reverse primer wR:GGGAGCATTTTCATGAGTACAACAA
Amplification mutant primer sequence:
Forward primer wmF:CAGAACAGACACTGGAACAAAATGA(is identical with wild-type F)
Reverse primer mR:GGTGGCCATACAAATCTAAGCCAG
Concrete treatment step is as follows:
(6) sample process
Get peripheral blood 2ml in the EDTA anticoagulant tube, gently put upside down mixing, 4 ℃ of preservations.
(2) DNA extraction
Get 200 μ L anticoagulations and extract DNA with Q IAmp DNA Blood Mini Kit test kit.Adopt the running gel imaging that DNA purity and concentration are detected;
(3) configuration of reaction system
Add following reaction system (PCR reagent is bought from TaKaRa) in the air in each reaction:
(4) increase by following program:
95℃3min
94 ℃ of 30sec → (50-60 ℃) all right 30sec → 72 ℃ 30sec, 40 circulations
72℃5min→4℃
(5) electrophoresis detection amplification
Join 2% sepharose, get 9 μ L(3) loading behind the product of amplification and the 10 times of concentration sample-loading buffer mixings, 120V electrophoresis 30min, ultraviolet judgement PCR result.When fragment deletion, can amplify the fragment of a 261bp; When this fragment does not lack, can amplify the fragment of a 365bp; If 261bp and 365bp product exist simultaneously, this sample is heterozygote.
This technology two different products that increase simultaneously in same reaction system are identified the sample genotype.And expanding fragment length is moderate, and the amplification condition wide scope uses instrument simple.
Compare with other method, the technology of the present invention has high specific, highly sensitive and advantage conveniently; And flux is higher, and the single cost is lower!
The description of above preferred embodiment makes those skilled in the art can make or use the present invention.The various modifications of these embodiment are apparent for a person skilled in the art, and the General Principle of definition can be applied among other embodiment and do not deviate from the spirit or scope of the present invention here.Therefore, the present invention is not limited to shown here embodiment, and will meet the most wide in range scope consistent with the principle that discloses and novel feature here.
Claims (7)
1. a method that detects the BCL2L11 gene pleiomorphism is characterized in that, it comprises following performing step:
Designed two pairs of primers, mutant primer and wild-type primer, pair of primers are crossed over the disappearance zone of 2903bp, when this fragment does not lack, and experiment condition limited reactions, the fragment of the length like this that can not increase; When this fragment deletion, can amplify the fragment of a 261bp;
Pair of primers when this fragment does not lack, can amplify the fragment of a 365bp in the disappearance zone of 2903bp in addition.
2. method according to claim 1, it is characterized in that: the wild-type primer sequence of described amplification is:
Forward primer wmF:CAGAACAGACACTGGAACAAAATGA
Reverse primer wR:GGGAGCATTTTCATGAGTACAACAA.
3. method according to claim 1, it is characterized in that: the mutant primer sequence of described amplification is:
Forward primer wmF:CAGAACAGACACTGGAACAAAATGA(is identical with wild-type F)
Reverse primer mR:GGTGGCCATACAAATCTAAGCCAG.
4. method according to claim 1, it is characterized in that: described concrete treatment step is as follows:
(1) sample process is got peripheral blood 2ml in the EDTA anticoagulant tube, gently puts upside down mixing, 4 ℃ of preservations;
(2) DNA extraction is got 200 μ L anticoagulations and is extracted DNA with test kit, adopts the running gel imaging that DNA purity and concentration are detected;
(3) configuration of reaction system is in the certain reaction system of the aerial adding of each reaction;
(4) increase by certain program;
(5) electrophoresis detection amplification.
5. method according to claim 4, it is characterized in that: described reaction system is as follows:
。
6. method according to claim 4 is characterized in that: describedly increase by following program:
95℃3min
94 ℃ of 30sec → (50-60 ℃) all right 30sec → 72 ℃ 30sec, 40 circulations
72℃5min→4℃。
7. method according to claim 4, it is characterized in that: described electrophoresis detection amplification step is as follows: join 2% sepharose, get 9 μ L(3) loading behind the product of amplification and the 10 times of concentration sample-loading buffer mixings, 120V electrophoresis 30min, ultraviolet judgement PCR result; When fragment deletion, can amplify the fragment of a 261bp; When this fragment does not lack, can amplify the fragment of a 365bp; If 261bp and 365bp product exist simultaneously, this sample is heterozygote.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210408118.9A CN102912022B (en) | 2012-10-24 | 2012-10-24 | Method for detecting polymorphism of BCL2L11 gene |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210408118.9A CN102912022B (en) | 2012-10-24 | 2012-10-24 | Method for detecting polymorphism of BCL2L11 gene |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102912022A true CN102912022A (en) | 2013-02-06 |
| CN102912022B CN102912022B (en) | 2014-05-28 |
Family
ID=47610597
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210408118.9A Expired - Fee Related CN102912022B (en) | 2012-10-24 | 2012-10-24 | Method for detecting polymorphism of BCL2L11 gene |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102912022B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112391465A (en) * | 2019-08-19 | 2021-02-23 | 宁波爱她基因科技有限公司 | Multi-gene mutation detection kit for lung cancer |
-
2012
- 2012-10-24 CN CN201210408118.9A patent/CN102912022B/en not_active Expired - Fee Related
Non-Patent Citations (1)
| Title |
|---|
| KING PAN NG ET AL: "A comon BIM deletion polymorphism mediates intrinsic resistance and inferior responses to tyrosine kinase inhibititors in cancer", 《NATURE MEDICINE》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112391465A (en) * | 2019-08-19 | 2021-02-23 | 宁波爱她基因科技有限公司 | Multi-gene mutation detection kit for lung cancer |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102912022B (en) | 2014-05-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Xu et al. | Circulating tumor DNA identified by targeted sequencing in advanced-stage non-small cell lung cancer patients | |
| CN108048531B (en) | Ultra-blocking fluorescent quantitative PCR method for detecting rare mutation with high sensitivity | |
| JP7191140B2 (en) | TERT promoter mutations in a subset of gliomas and tumors | |
| Borczuk et al. | Genome-wide analysis of abdominal and pleural malignant mesothelioma with DNA arrays reveals both common and distinct regions of copy number alteration | |
| Hostetler et al. | Independent origin and global distribution of distinct Plasmodium vivax Duffy binding protein gene duplications | |
| CN104293913B (en) | Multipoint mutation single tube method for quick and test kit | |
| CN104450948B (en) | Method for detecting cancer, kit and its application | |
| CN110343748B (en) | Method for analyzing tumor mutation load based on high-throughput targeted sequencing | |
| CN104774962A (en) | Kit for detecting BRAF gene mutation and detecting method thereof | |
| CN104561250A (en) | Method and primer for detecting imatinib targeted medication gene | |
| Mariani et al. | Concordant analysis of KRAS status in primary colon carcinoma and matched metastasis | |
| Youssef et al. | Hotspot mutations detectable by next-generation sequencing in exhaled breath condensates from patients with lung cancer | |
| CN104017887A (en) | Primer pair as well as probe and kit for detecting human BRAF gene mutation | |
| JP2023505031A (en) | Methods and compositions for cancer analysis | |
| CN105803076A (en) | Specific primer and probe for detecting T790M locus of EGFR gene | |
| CN104480215B (en) | A kind of gene association detection method and test kit | |
| CN102912022B (en) | Method for detecting polymorphism of BCL2L11 gene | |
| CN103103284B (en) | Kit for quantitatively detecting IKZF1 gene mutation | |
| CN111118119A (en) | Method for detecting target mutation by carrying out retardation substitution amplification enrichment based on blocker introducing extra base mismatch | |
| CN107385066A (en) | Detect primer pair, kit, detection method and the application of EGFR genetic mutation | |
| Setty et al. | Sensitive determination of BRAF copy number in clinical samples by pyrosequencing | |
| CN109554475A (en) | Gene mutation/fusion combination and kit for Lung neoplasm malignant and benign lesion | |
| Mäki-Nevala et al. | Concordant results of epidermal growth factor receptor mutation detection by real-time polymerase chain reaction and ion torrent technology in non-small cell lung cancer | |
| Hernández et al. | Novel multiplex RT-PCR/RFLP diagnostic test to differentiate low-from high-pathogenic strains and to detect reassortant infectious bursal disease virus | |
| CN103898232B (en) | A kind of method detecting low abundant species and change |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20201111 Address after: 274000 No. 2888 Zhonghua West Road, hi tech Zone, Shandong, Heze Patentee after: GUOJIUTANG SHANDONG DONKEY-HIDE GELATIN Co.,Ltd. Address before: Room 209d, 668 SHANGDA Road, Baoshan District, Shanghai Patentee before: SHANGHAI SHINES PHARMACEUTICAL Co.,Ltd. |
|
| TR01 | Transfer of patent right | ||
| CP01 | Change in the name or title of a patent holder |
Address after: No.2888, Zhonghua West Road, hi tech Zone, Heze City, Shandong Province 274000 Patentee after: Shandong Guojiutang Pharmaceutical Group Co.,Ltd. Address before: No.2888, Zhonghua West Road, hi tech Zone, Heze City, Shandong Province 274000 Patentee before: GUOJIUTANG SHANDONG DONKEY-HIDE GELATIN CO.,LTD. |
|
| CP01 | Change in the name or title of a patent holder | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140528 |