CN102392047B - Bicistronic mRNA (messenger ribonucleic acid) expression vector suitable for cells of mammals and application thereof - Google Patents
Bicistronic mRNA (messenger ribonucleic acid) expression vector suitable for cells of mammals and application thereof Download PDFInfo
- Publication number
- CN102392047B CN102392047B CN201110355135.6A CN201110355135A CN102392047B CN 102392047 B CN102392047 B CN 102392047B CN 201110355135 A CN201110355135 A CN 201110355135A CN 102392047 B CN102392047 B CN 102392047B
- Authority
- CN
- China
- Prior art keywords
- expression vector
- restriction enzyme
- open reading
- reading frame
- enzyme site
- 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
- 239000013604 expression vector Substances 0.000 title claims abstract description 41
- 241000124008 Mammalia Species 0.000 title abstract 2
- 229920002477 rna polymer Polymers 0.000 title abstract 2
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 43
- 108700026244 Open Reading Frames Proteins 0.000 claims abstract description 32
- 230000014509 gene expression Effects 0.000 claims abstract description 32
- 241000701022 Cytomegalovirus Species 0.000 claims abstract description 23
- 241000829100 Macaca mulatta polyomavirus 1 Species 0.000 claims abstract description 16
- 230000005030 transcription termination Effects 0.000 claims abstract description 10
- 230000035897 transcription Effects 0.000 claims abstract description 9
- 238000013518 transcription Methods 0.000 claims abstract description 9
- 108091008146 restriction endonucleases Proteins 0.000 claims description 63
- 239000013612 plasmid Substances 0.000 claims description 27
- 210000004962 mammalian cell Anatomy 0.000 claims description 24
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 14
- 108010030074 endodeoxyribonuclease MluI Proteins 0.000 claims description 10
- 108700007698 Genetic Terminator Regions Proteins 0.000 claims description 7
- 210000004978 chinese hamster ovary cell Anatomy 0.000 claims description 4
- 230000009466 transformation Effects 0.000 claims description 3
- 108010022394 Threonine synthase Proteins 0.000 claims 1
- 238000012216 screening Methods 0.000 abstract description 9
- 238000004519 manufacturing process Methods 0.000 abstract description 5
- 230000004544 DNA amplification Effects 0.000 abstract description 3
- 108010006025 bovine growth hormone Proteins 0.000 abstract 2
- 239000003623 enhancer Substances 0.000 abstract 2
- 241000700605 Viruses Species 0.000 abstract 1
- 239000003550 marker Substances 0.000 abstract 1
- 210000004027 cell Anatomy 0.000 description 36
- 238000006243 chemical reaction Methods 0.000 description 32
- 239000000047 product Substances 0.000 description 29
- 230000003321 amplification Effects 0.000 description 18
- 238000003199 nucleic acid amplification method Methods 0.000 description 18
- 102100040247 Tumor necrosis factor Human genes 0.000 description 17
- 239000012634 fragment Substances 0.000 description 17
- 239000003292 glue Substances 0.000 description 13
- 102000004169 proteins and genes Human genes 0.000 description 13
- 108020004414 DNA Proteins 0.000 description 11
- 238000000246 agarose gel electrophoresis Methods 0.000 description 11
- 230000029087 digestion Effects 0.000 description 11
- 238000000034 method Methods 0.000 description 11
- 238000012360 testing method Methods 0.000 description 11
- 238000013467 fragmentation Methods 0.000 description 10
- 238000006062 fragmentation reaction Methods 0.000 description 10
- 238000013461 design Methods 0.000 description 8
- 238000011084 recovery Methods 0.000 description 8
- 238000001890 transfection Methods 0.000 description 8
- 238000011144 upstream manufacturing Methods 0.000 description 8
- FBOZXECLQNJBKD-ZDUSSCGKSA-N L-methotrexate Chemical compound C=1N=C2N=C(N)N=C(N)C2=NC=1CN(C)C1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 FBOZXECLQNJBKD-ZDUSSCGKSA-N 0.000 description 7
- 102000005396 glutamine synthetase Human genes 0.000 description 7
- 108020002326 glutamine synthetase Proteins 0.000 description 7
- 229960000485 methotrexate Drugs 0.000 description 7
- 102100024746 Dihydrofolate reductase Human genes 0.000 description 6
- 108020001096 dihydrofolate reductase Proteins 0.000 description 6
- 235000015097 nutrients Nutrition 0.000 description 6
- 238000005406 washing Methods 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 5
- 108090000790 Enzymes Proteins 0.000 description 5
- 238000012408 PCR amplification Methods 0.000 description 5
- 239000002299 complementary DNA Substances 0.000 description 5
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 239000000243 solution Substances 0.000 description 5
- 244000309466 calf Species 0.000 description 4
- 108020004999 messenger RNA Proteins 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 3
- 241000699802 Cricetulus griseus Species 0.000 description 3
- 238000005520 cutting process Methods 0.000 description 3
- 239000013613 expression plasmid Substances 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 210000001672 ovary Anatomy 0.000 description 3
- 238000002360 preparation method Methods 0.000 description 3
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- 108700007696 Tetrahydrofolate Dehydrogenase Proteins 0.000 description 2
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 2
- 101150099105 alien gene Proteins 0.000 description 2
- 230000001079 digestive effect Effects 0.000 description 2
- 102000004419 dihydrofolate reductase Human genes 0.000 description 2
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 230000013595 glycosylation Effects 0.000 description 2
- 238000006206 glycosylation reaction Methods 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 210000005259 peripheral blood Anatomy 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 206010039073 rheumatoid arthritis Diseases 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 101100230376 Acetivibrio thermocellus (strain ATCC 27405 / DSM 1237 / JCM 9322 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372) celI gene Proteins 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 1
- 238000001712 DNA sequencing Methods 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 101150074355 GS gene Proteins 0.000 description 1
- 108700028146 Genetic Enhancer Elements Proteins 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 101001091385 Homo sapiens Kallikrein-6 Proteins 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102100034866 Kallikrein-6 Human genes 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 108091061960 Naked DNA Proteins 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 108091034057 RNA (poly(A)) Proteins 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 150000003863 ammonium salts Chemical class 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 235000011089 carbon dioxide Nutrition 0.000 description 1
- 238000006555 catalytic reaction Methods 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000012790 confirmation Methods 0.000 description 1
- 210000004748 cultured cell Anatomy 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000001186 cumulative effect Effects 0.000 description 1
- 238000013016 damping Methods 0.000 description 1
- 230000009849 deactivation Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 229930195712 glutamate Natural products 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-M glutaminate Chemical compound [O-]C(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-M 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- -1 nitrite ions Chemical class 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 239000012679 serum free medium Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
Description
Claims (14)
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201110355135.6A CN102392047B (en) | 2011-11-11 | 2011-11-11 | Bicistronic mRNA (messenger ribonucleic acid) expression vector suitable for cells of mammals and application thereof |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201110355135.6A CN102392047B (en) | 2011-11-11 | 2011-11-11 | Bicistronic mRNA (messenger ribonucleic acid) expression vector suitable for cells of mammals and application thereof |
Publications (2)
Publication Number | Publication Date |
---|---|
CN102392047A CN102392047A (en) | 2012-03-28 |
CN102392047B true CN102392047B (en) | 2014-01-01 |
Family
ID=45859427
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201110355135.6A Active CN102392047B (en) | 2011-11-11 | 2011-11-11 | Bicistronic mRNA (messenger ribonucleic acid) expression vector suitable for cells of mammals and application thereof |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN102392047B (en) |
Families Citing this family (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DK2711426T3 (en) * | 2012-09-24 | 2015-07-13 | Lonza Biologics Plc | Expression vectors comprising chimeric cytomegalovirus promoter and enhancer sequences |
EP3683314A1 (en) | 2013-10-07 | 2020-07-22 | Prestige Biopharma Pte. Ltd. | Bicistronic expression vector for antibody expression and method for producing antibody using same |
CN109957578A (en) * | 2017-12-22 | 2019-07-02 | 拜西欧斯(北京)生物技术有限公司 | Double gene expression vector and its preparation method and application |
CN109913484A (en) * | 2019-01-14 | 2019-06-21 | 陕西理工大学 | A kind of two-way expression carrier T with and its preparation method and application |
Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101705244A (en) * | 2009-11-23 | 2010-05-12 | 上海凯茂生物医药有限公司 | Animal cell bicistronic high-efficiency expression vector |
Family Cites Families (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101233155B (en) * | 2004-03-19 | 2013-06-19 | 英克隆有限责任公司 | Human anti-epidermal growth factor receptor antibody |
-
2011
- 2011-11-11 CN CN201110355135.6A patent/CN102392047B/en active Active
Patent Citations (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101705244A (en) * | 2009-11-23 | 2010-05-12 | 上海凯茂生物医药有限公司 | Animal cell bicistronic high-efficiency expression vector |
Non-Patent Citations (4)
Title |
---|
哺乳动物细胞高效表达系统研究进展;张国强;《生物技术通讯》;20050131;第16卷(第1期);56-59 * |
基于FMDV IRES的双顺反子载体的构建及体外表达分析;郑海学;《中国生物工程杂志》;20071231;第27卷(第8期);29-33 * |
张国强.哺乳动物细胞高效表达系统研究进展.《生物技术通讯》.2005,第16卷(第1期),56-59. |
郑海学.基于FMDV IRES的双顺反子载体的构建及体外表达分析.《中国生物工程杂志》.2007,第27卷(第8期),29-33. |
Also Published As
Publication number | Publication date |
---|---|
CN102392047A (en) | 2012-03-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN107828738A (en) | A kind of dnmt rna deficiency Chinese hamster ovary celI system and preparation method and application | |
CN102165060B (en) | Novel regulatory elements | |
CN101903523B (en) | Mammalian expression vector | |
CN102392047B (en) | Bicistronic mRNA (messenger ribonucleic acid) expression vector suitable for cells of mammals and application thereof | |
US9260721B2 (en) | Expression vector and methods of producing high levels of proteins | |
CN111944841A (en) | Arginine deiminase secreted and expressed by pichia pastoris and construction method | |
CN107868781A (en) | Artificial synthesized MAR fragments, expression vector, expression system and its application | |
CN107760650A (en) | A kind of Chinese hamster ovary celI of transformation and application thereof | |
CN103298925A (en) | Methods for culturing human myeloid leukaemia cells and cells derived therefrom | |
CN101341252B (en) | Protein expression in rodent cells | |
Kitchin et al. | Alteration of hybridoma viability and antibody secretion in transfectomas with inducible overexpression of protein disulfide isomerase | |
CN101415824B (en) | Auto-inducible sodium phosphate symporter promoter from pichia pastoris and method for producing recombinant protein using it | |
CN110343699A (en) | For improving regulating and controlling sequence, expression vector, expression system and its application of mammalian cell transgene expression | |
CN104195173B (en) | Glutamine synthelase efficient expression vector with double expression boxes | |
CN101163792A (en) | Mammalian expression vector comprising the mcmv promoter and first intron of hcmv major immediate early gene | |
MX2013010872A (en) | Regulated gene expression systems and constructs thereof. | |
CN115838713A (en) | Protease and application thereof in L-carnosine synthesis | |
CN110343718A (en) | A kind of fibrocyte expression vector of efficient stable, expression system and preparation method thereof, application | |
CN101988049A (en) | Genetically modified cell of coexpression mouse membranous type interleukin 15 and retinoic acid early transcript 1 epsilon (Rae-1 epsilon) and preparation method thereof | |
CN103483423B (en) | Artificially synthesized signal peptide and application thereof | |
CN1244696C (en) | Highly effective expressing method of interleukin-12 | |
CN104975018B (en) | A kind of novel enhanced is sub and its applies | |
CN116622702A (en) | Novel artificially designed bacillus subtilis terminator and application thereof | |
CN114807145B (en) | Leader sequence for improving cap-independent translation efficiency and application thereof | |
CN106177906B (en) | The application of Ddb1 albumen |
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: 20181214 Address after: Room 3302, Room 3, 102 Location, Building 797, Puxing Road, Minhang District, Shanghai, 20114 Patentee after: Shanghai Chenjian Biotechnology Co.,Ltd. Address before: 215125 Room 111, A3 Building, 218 Xinghu Street, Suzhou Industrial Park, Jiangsu Province Patentee before: Suzhou Industrial Park Chenjian Antibody Group Pharmaceutical Development Co.,Ltd. |
|
TR01 | Transfer of patent right | ||
TR01 | Transfer of patent right |
Effective date of registration: 20210902 Address after: 518118 unit 1505, block a, innovation Plaza, No. 2007, Pingshan street, Pingshan District, Shenzhen, Guangdong Patentee after: Yourui Biomedical Technology (Shenzhen) Co.,Ltd. Address before: Room 3302, Room 3, 102 Location, Building 797, Puxing Road, Minhang District, Shanghai, 20114 Patentee before: Shanghai Chenjian Biotechnology Co.,Ltd. |
|
TR01 | Transfer of patent right | ||
PP01 | Preservation of patent right |
Effective date of registration: 20230522 Granted publication date: 20140101 |
|
PP01 | Preservation of patent right | ||
PD01 | Discharge of preservation of patent |
Date of cancellation: 20230815 Granted publication date: 20140101 |
|
PD01 | Discharge of preservation of patent | ||
CP03 | Change of name, title or address |
Address after: 518118 unit 1505, block a, innovation Plaza, No. 2007, Pingshan street, Pingshan District, Shenzhen, Guangdong Patentee after: Youjian Biopharmaceutical Technology (Shenzhen) Co.,Ltd. Country or region after: China Address before: 518118 unit 1505, block a, innovation Plaza, No. 2007, Pingshan street, Pingshan District, Shenzhen, Guangdong Patentee before: Yourui Biomedical Technology (Shenzhen) Co.,Ltd. Country or region before: China |
|
CP03 | Change of name, title or address |