CN114292853A - SUMO (small cell-associated protein) lysis-promoting expression tag and application thereof, SUMO lysis-promoting expression protein, recombinant strain and preparation method of protein - Google Patents
SUMO (small cell-associated protein) lysis-promoting expression tag and application thereof, SUMO lysis-promoting expression protein, recombinant strain and preparation method of protein Download PDFInfo
- Publication number
- CN114292853A CN114292853A CN202111446975.3A CN202111446975A CN114292853A CN 114292853 A CN114292853 A CN 114292853A CN 202111446975 A CN202111446975 A CN 202111446975A CN 114292853 A CN114292853 A CN 114292853A
- Authority
- CN
- China
- Prior art keywords
- sumo
- protein
- expression
- solubilizing
- tag
- 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
- 230000014509 gene expression Effects 0.000 title claims abstract description 124
- 108090000623 proteins and genes Proteins 0.000 title claims abstract description 87
- 102000004169 proteins and genes Human genes 0.000 title claims abstract description 83
- 238000002360 preparation method Methods 0.000 title claims abstract description 26
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 title description 3
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 title description 3
- 230000003381 solubilizing effect Effects 0.000 claims abstract description 27
- 102000051619 SUMO-1 Human genes 0.000 claims abstract description 16
- 238000001976 enzyme digestion Methods 0.000 claims abstract description 15
- 239000013604 expression vector Substances 0.000 claims abstract description 14
- 101710194807 Protective antigen Proteins 0.000 claims abstract description 13
- 230000009465 prokaryotic expression Effects 0.000 claims abstract description 9
- 239000002773 nucleotide Substances 0.000 claims abstract description 7
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 7
- 101000879203 Caenorhabditis elegans Small ubiquitin-related modifier Proteins 0.000 claims abstract 15
- 208000012153 swine disease Diseases 0.000 claims description 8
- 102000036639 antigens Human genes 0.000 claims description 6
- 108091007433 antigens Proteins 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 238000012408 PCR amplification Methods 0.000 claims description 5
- 239000000427 antigen Substances 0.000 claims description 5
- 238000006243 chemical reaction Methods 0.000 claims description 5
- 101710204837 Envelope small membrane protein Proteins 0.000 claims description 4
- 101710145006 Lysis protein Proteins 0.000 claims description 4
- 101710141454 Nucleoprotein Proteins 0.000 claims description 4
- 241000711484 Transmissible gastroenteritis virus Species 0.000 claims description 4
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 claims description 3
- 241000202347 Porcine circovirus Species 0.000 claims description 3
- 241000701076 Macacine alphaherpesvirus 1 Species 0.000 claims 1
- 125000003275 alpha amino acid group Chemical group 0.000 claims 1
- 206010014599 encephalitis Diseases 0.000 claims 1
- 238000009472 formulation Methods 0.000 claims 1
- 239000000203 mixture Substances 0.000 claims 1
- 229940031626 subunit vaccine Drugs 0.000 abstract description 15
- 238000004519 manufacturing process Methods 0.000 abstract description 12
- 241000282887 Suidae Species 0.000 abstract description 9
- 238000007086 side reaction Methods 0.000 abstract description 7
- 108020004705 Codon Proteins 0.000 abstract description 6
- 230000003053 immunization Effects 0.000 abstract description 5
- 238000002649 immunization Methods 0.000 abstract description 5
- 238000010353 genetic engineering Methods 0.000 abstract description 3
- 238000005457 optimization Methods 0.000 abstract 1
- 108010043401 Small Ubiquitin-Related Modifier Proteins Proteins 0.000 description 77
- 102000002669 Small Ubiquitin-Related Modifier Proteins Human genes 0.000 description 64
- 241000282898 Sus scrofa Species 0.000 description 28
- 102000037865 fusion proteins Human genes 0.000 description 19
- 108020001507 fusion proteins Proteins 0.000 description 19
- 241000588724 Escherichia coli Species 0.000 description 17
- 239000000047 product Substances 0.000 description 13
- 239000006228 supernatant Substances 0.000 description 10
- 238000005119 centrifugation Methods 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 8
- 238000001962 electrophoresis Methods 0.000 description 8
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 8
- 239000013598 vector Substances 0.000 description 7
- 239000013612 plasmid Substances 0.000 description 6
- 238000004090 dissolution Methods 0.000 description 5
- 230000001737 promoting effect Effects 0.000 description 5
- 238000003259 recombinant expression Methods 0.000 description 5
- 230000007928 solubilization Effects 0.000 description 5
- 238000005063 solubilization Methods 0.000 description 5
- 108700038981 SUMO-1 Proteins 0.000 description 4
- 238000000246 agarose gel electrophoresis Methods 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 239000012634 fragment Substances 0.000 description 4
- 230000004927 fusion Effects 0.000 description 4
- 239000000499 gel Substances 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 108020004414 DNA Proteins 0.000 description 3
- 241000710842 Japanese encephalitis virus Species 0.000 description 3
- 108010006519 Molecular Chaperones Proteins 0.000 description 3
- 102000005431 Molecular Chaperones Human genes 0.000 description 3
- 229930027917 kanamycin Natural products 0.000 description 3
- SBUJHOSQTJFQJX-NOAMYHISSA-N kanamycin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N SBUJHOSQTJFQJX-NOAMYHISSA-N 0.000 description 3
- 229960000318 kanamycin Drugs 0.000 description 3
- 229930182823 kanamycin A Natural products 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 108091008146 restriction endonucleases Proteins 0.000 description 3
- 101000708016 Caenorhabditis elegans Sentrin-specific protease Proteins 0.000 description 2
- 102000012410 DNA Ligases Human genes 0.000 description 2
- 108010061982 DNA Ligases Proteins 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 150000001413 amino acids Chemical group 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 210000003000 inclusion body Anatomy 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000006911 nucleation Effects 0.000 description 2
- 238000010899 nucleation Methods 0.000 description 2
- 230000001323 posttranslational effect Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- BLGHHPHXVJWCNK-GUBZILKMSA-N Ala-Gln-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O BLGHHPHXVJWCNK-GUBZILKMSA-N 0.000 description 1
- WMYJZJRILUVVRG-WDSKDSINSA-N Ala-Gly-Gln Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCC(N)=O WMYJZJRILUVVRG-WDSKDSINSA-N 0.000 description 1
- CVKOQHYVDVYJSI-QTKMDUPCSA-N Arg-His-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CCCN=C(N)N)N)O CVKOQHYVDVYJSI-QTKMDUPCSA-N 0.000 description 1
- FKQITMVNILRUCQ-IHRRRGAJSA-N Arg-Phe-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O FKQITMVNILRUCQ-IHRRRGAJSA-N 0.000 description 1
- GFFRWIJAFFMQGM-NUMRIWBASA-N Asn-Glu-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GFFRWIJAFFMQGM-NUMRIWBASA-N 0.000 description 1
- SNDBKTFJWVEVPO-WHFBIAKZSA-N Asp-Gly-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(O)=O SNDBKTFJWVEVPO-WHFBIAKZSA-N 0.000 description 1
- PDIYGFYAMZZFCW-JIOCBJNQSA-N Asp-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC(=O)O)N)O PDIYGFYAMZZFCW-JIOCBJNQSA-N 0.000 description 1
- 241001533384 Circovirus Species 0.000 description 1
- 241000588914 Enterobacter Species 0.000 description 1
- 241000206602 Eukaryota Species 0.000 description 1
- OREPWMPAUWIIAM-ZPFDUUQYSA-N Gln-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CCC(=O)N)N OREPWMPAUWIIAM-ZPFDUUQYSA-N 0.000 description 1
- ININBLZFFVOQIO-JHEQGTHGSA-N Gln-Thr-Gly Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CCC(=O)N)N)O ININBLZFFVOQIO-JHEQGTHGSA-N 0.000 description 1
- RCCDHXSRMWCOOY-GUBZILKMSA-N Glu-Arg-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O RCCDHXSRMWCOOY-GUBZILKMSA-N 0.000 description 1
- GLWXKFRTOHKGIT-ACZMJKKPSA-N Glu-Asn-Asn Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O GLWXKFRTOHKGIT-ACZMJKKPSA-N 0.000 description 1
- SBCYJMOOHUDWDA-NUMRIWBASA-N Glu-Asp-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SBCYJMOOHUDWDA-NUMRIWBASA-N 0.000 description 1
- FMBWLLMUPXTXFC-SDDRHHMPSA-N Glu-Lys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(=O)O)N)C(=O)O FMBWLLMUPXTXFC-SDDRHHMPSA-N 0.000 description 1
- JHSRJMUJOGLIHK-GUBZILKMSA-N Glu-Met-Glu Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CCC(=O)O)N JHSRJMUJOGLIHK-GUBZILKMSA-N 0.000 description 1
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 1
- IZVICCORZOSGPT-JSGCOSHPSA-N Gly-Val-Tyr Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O IZVICCORZOSGPT-JSGCOSHPSA-N 0.000 description 1
- VJJSDSNFXCWCEJ-DJFWLOJKSA-N His-Ile-Asn Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O VJJSDSNFXCWCEJ-DJFWLOJKSA-N 0.000 description 1
- VZIFYHYNQDIPLI-HJWJTTGWSA-N Ile-Arg-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N VZIFYHYNQDIPLI-HJWJTTGWSA-N 0.000 description 1
- LLZLRXBTOOFODM-QSFUFRPTSA-N Ile-Asp-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](C(C)C)C(=O)O)N LLZLRXBTOOFODM-QSFUFRPTSA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- LZHJZLHSRGWBBE-IHRRRGAJSA-N Leu-Lys-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O LZHJZLHSRGWBBE-IHRRRGAJSA-N 0.000 description 1
- VHXMZJGOKIMETG-CQDKDKBSSA-N Lys-Ala-Tyr Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)NC(=O)[C@H](CCCCN)N VHXMZJGOKIMETG-CQDKDKBSSA-N 0.000 description 1
- GCMWRRQAKQXDED-IUCAKERBSA-N Lys-Glu-Gly Chemical compound [NH3+]CCCC[C@H]([NH3+])C(=O)N[C@@H](CCC([O-])=O)C(=O)NCC([O-])=O GCMWRRQAKQXDED-IUCAKERBSA-N 0.000 description 1
- QOJDBRUCOXQSSK-AJNGGQMLSA-N Lys-Ile-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(O)=O QOJDBRUCOXQSSK-AJNGGQMLSA-N 0.000 description 1
- ORVFEGYUJITPGI-IHRRRGAJSA-N Lys-Leu-Met Chemical compound CSCC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCCCN ORVFEGYUJITPGI-IHRRRGAJSA-N 0.000 description 1
- VHGIWFGJIHTASW-FXQIFTODSA-N Met-Ala-Asp Chemical compound CSCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O VHGIWFGJIHTASW-FXQIFTODSA-N 0.000 description 1
- IIPHCNKHEZYSNE-DCAQKATOSA-N Met-Arg-Gln Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O IIPHCNKHEZYSNE-DCAQKATOSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- IILUKIJNFMUBNF-IHRRRGAJSA-N Phe-Gln-Gln Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O IILUKIJNFMUBNF-IHRRRGAJSA-N 0.000 description 1
- FKYKZHOKDOPHSA-DCAQKATOSA-N Pro-Leu-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O FKYKZHOKDOPHSA-DCAQKATOSA-N 0.000 description 1
- 102000044159 Ubiquitin Human genes 0.000 description 1
- 108090000848 Ubiquitin Proteins 0.000 description 1
- XPKCFQZDQGVJCX-RHYQMDGZSA-N Val-Lys-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C(C)C)N)O XPKCFQZDQGVJCX-RHYQMDGZSA-N 0.000 description 1
- VVIZITNVZUAEMI-DLOVCJGASA-N Val-Val-Gln Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCC(N)=O VVIZITNVZUAEMI-DLOVCJGASA-N 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 108010047857 aspartylglycine Proteins 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 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 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 230000002101 lytic effect Effects 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 108010072637 phenylalanyl-arginyl-phenylalanine Proteins 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000004481 post-translational protein modification Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 108010048397 seryl-lysyl-leucine Proteins 0.000 description 1
- 108010071207 serylmethionine Proteins 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 229960005486 vaccine Drugs 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
Images
Landscapes
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention provides a SUMO (soluble-associated protein) dissolving-promoting expression label and application thereof, a SUMO dissolving-promoting expression protein, a recombinant strain and a preparation method of the protein, and relates to the technical field of genetic engineering, wherein the nucleotide sequence of the SUMO dissolving-promoting expression label is shown as SEQ ID NO: 1, the SUMO solubilizing expression label provided by the invention is derived from a pig SUMO gene sequence after codon optimization, is derived from a host pig, and is fused with a prokaryotic expression vector to perform protein expression, so that correct folding of target protein can be better promoted, and the soluble expression quantity of foreign protein is improved. The SUMO solubilizing expression tag provided by the invention is derived from a host pig, when the SUMO solubilizing expression tag is applied to preparation of a subunit vaccine for pigs, side reactions after immunization can not be caused, enzyme digestion of the SUMO solubilizing expression tag is not needed, the solubility and the expression quantity of a protective antigen can be improved, the preparation process of the subunit vaccine for pigs can be simplified, the production cost is reduced, and the production efficiency is improved.
Description
Technical Field
The invention relates to the technical field of genetic engineering, and relates to a SUMO (soluble protein synthase) solubilization expression label, application thereof, a SUMO solubilization expression protein, a recombinant strain and a preparation method of the protein.
Background
Because a prokaryotic expression system lacks a proper post-translational processing mechanism, an insoluble inclusion body is formed due to incorrect folding in the process of expressing exogenous protein by using escherichia coli as host bacteria, and the high soluble expression quantity is difficult to obtain.
Disclosure of Invention
The invention mainly aims to provide a SUMO (soluble protein-associated protein) solubilization expression label and application thereof, a SUMO solubilization expression protein, a recombinant strain and a preparation method of the protein, and aims to provide the SUMO solubilization expression label capable of improving the solubility and the expression quantity of the protein.
In order to achieve the technical purpose, the invention provides a fused SUMO dissolving promotion expression label, wherein the nucleotide sequence of the SUMO dissolving promotion expression label is shown as SEQ ID NO: 1 is shown.
The invention further provides a SUMO (soluble protein) expression promoting protein, wherein the amino acid sequence of the SUMO expression promoting protein is shown as SEQ ID NO: 2, respectively.
The invention further provides a recombinant strain comprising the SUMO-facilitated expression tag as described above.
Optionally, the recombinant strain comprises escherichia coli.
The invention further provides a preparation method of the protein, which comprises the following steps:
performing PCR amplification by using the SUMO solubilizing expression label as a template to obtain an SUMO gene;
inserting the SUMO gene into a prokaryotic expression vector through enzyme digestion reaction to obtain an expression system;
expressing the protein according to the expression system to obtain the protein;
wherein, the nucleotide sequence of the SUMO solubilizing and expressing label is shown as SEQ ID NO: 1 is shown.
Optionally, the protein comprises a swine disease protective antigen.
Optionally, the swine disease protective antigen comprises swine Japanese encephalitis virus E protein, swine transmissible gastroenteritis virus N protein, and porcine circovirus type 3cap protein.
The invention further provides an application of the SUMO solubilizing expression label in preparing an antigen preparation.
The SUMO dissolving promotion expression label provided by the invention is derived from a codon optimized pig SUMO gene sequence and a host pig, and the base sequence of the SUMO dissolving promotion expression label is shown as SEQ ID No: 1, the porcine small ubiquitin-like modified protein SUMO with optimized codons is used as a solubility-promoting expression label, and is fused with a prokaryotic expression vector for protein expression, so that correct folding of target protein can be better promoted, and the soluble expression quantity of foreign protein is improved. The SUMO solubilizing expression tag provided by the invention is derived from a host pig, when the SUMO solubilizing expression tag is applied to preparation of a subunit vaccine for pigs, side reactions after immunization can not be caused, enzyme digestion of the SUMO solubilizing expression tag is not needed, the solubility and the expression quantity of a protective antigen can be improved, the preparation process of the subunit vaccine for pigs can be simplified, the production cost is reduced, and the production efficiency is improved.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other related drawings can be obtained according to the drawings without creative efforts.
FIG. 1 is a diagram of agarose gel electrophoresis for recovery of SUMO gene gel obtained by PCR amplification of the SUMO facilitated digestion expression tag of example 1 of the present invention;
FIG. 2 is an SDS-PAGE electrophoresis chart of JEV-E, JEV-E-SUMO fusion protein and enzyme-cleaved JEV-E-SUMO fusion protein in example 2 of the present invention;
FIG. 3 is a SDS-PAGE electrophoresis chart of TGEV-N, TGEV-N-SUMO fusion protein and enzyme-digested TGEV-N-SUMO fusion protein in example 2 of the present invention;
FIG. 4 is an SDS-PAGE electrophoresis chart of PCV3cap, PCV3cap-SUMO fusion protein and enzyme-cleaved PCV3cap-SUMO fusion protein in example 2 of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
Because a prokaryotic expression system lacks a proper post-translational processing mechanism, an insoluble inclusion body is formed due to incorrect folding in the process of expressing exogenous protein by using escherichia coli as host bacteria, and the high soluble expression quantity is difficult to obtain.
In view of this, the present invention provides a SUMO solubility-promoting expression tag, and aims to provide a SUMO solubility-promoting expression tag capable of improving the solubility and expression level of a protein, and the SUMO solubility-promoting expression tag provided by the present invention has a nucleotide sequence shown in SEQ ID NO: 1 is shown.
The SUMO dissolving promotion expression label provided by the invention is derived from a codon optimized pig SUMO gene sequence and a host pig, and the base sequence of the SUMO dissolving promotion expression label is shown as SEQ ID No: 1, the porcine small ubiquitin-like modified protein SUMO with optimized codons is used as a solubility-promoting expression label, and is fused with a prokaryotic expression vector for protein expression, so that correct folding of target protein can be better promoted, and the soluble expression quantity of foreign protein is improved. The SUMO solubilizing expression tag is derived from a host pig, when the SUMO solubilizing expression tag is applied to preparation of the subunit vaccine for the pig, side reactions after immunization can not be caused, enzyme digestion of the SUMO solubilizing expression tag is not needed, the solubility and the expression quantity of a protective antigen can be improved, the preparation process of the subunit vaccine for the pig can be simplified, the production cost is reduced, and the production efficiency is improved.
The Small ubiquitin-like modified protein SUMO (Small ubiquitin-like modifier) is a molecular chaperone, has a structure similar to ubiquitin (beta-sheet is wound by an alpha-spiral spherical folding), conservatively exists in eukaryotes, and participates in various physiological processes such as protein posttranslational modification and the like. The SUMO protein has a highly hydrophobic core, and can provide nucleation sites for folding of fusion protein, so that the SUMO protein can be used as a molecular chaperone and a fusion label for improving stability and solubility of foreign protein, the tertiary structure of the SUMO protein can be completely identified by SUMO protease, the target protein after specific enzyme digestion has a natural N terminal, the activity of the target protein is ensured, and a SUMO expression system is obviously superior to a traditional expression system in terms of expression level and product solubility.
The SUMO genes are found in 5 types so far, and the SUMO dissolution promotion expression labels currently applied to the swine vaccines are mainly derived from human sources, and the swine source dissolution promotion labels are rarely applied. The SUMO solubilizing-promoting expression label is derived from a host pig, and is subjected to fusion expression with a swine disease protective antigen in escherichia coli, a swine-derived SUMO solubilizing-promoting expression label is not required to be removed after the fusion protein is expressed, a subunit vaccine for pigs can be directly prepared, and after a pig is immunized, a side reaction caused by the SUMO solubilizing-promoting expression label is avoided, so that the soluble expression of target protein can be promoted, and the effect of simplifying the preparation process of the subunit vaccine for pigs can be achieved.
The invention further provides a SUMO (soluble protein) expression promoting protein, wherein the amino acid sequence of the SUMO expression promoting protein is shown as SEQ ID NO: 2, respectively.
The SUMO pro-lytic expression protein provided by the invention can be expressed by a sequence shown as SEQ ID NO: 1, or other nucleotide sequences, and the SUMO-facilitated expression protein can provide nucleation sites for folding of the fusion protein, thereby serving as a molecular chaperone and a fusion tag for improving stability and solubility of foreign proteins, and the tertiary structure of the SUMO protein can be completely identified by SUMO protease, and the target protein after specific enzyme digestion has a natural N-terminal, so that the activity of the target protein is ensured, and the SUMO expression system is obviously superior to the conventional expression system in terms of expression level and product solubility.
The invention further provides a recombinant strain comprising the SUMO-facilitated expression tag as described above.
The recombinant strain provided by the invention is used for expressing the fusion protein consisting of the SUMO solubilizing expression protein and the target protein, preferably, the recombinant strain comprises escherichia coli, the genetic background of the escherichia coli is clear, a carrier receptor system is complete, the growth is rapid, the culture is simple, a recombinant is stable, and the recombinant strain is more suitable for the expression of prokaryotic genes.
The invention further provides a preparation method of the protein, which comprises the following steps:
s10, carrying out PCR amplification by taking the SUMO solubilizing expression label as a template to obtain the SUMO gene, wherein the nucleotide sequence of the SUMO solubilizing expression label is shown as SEQ ID NO: 1 is shown in the specification;
s20, inserting the SUMO gene into a prokaryotic expression vector through enzyme digestion reaction to obtain an expression system;
and S30, expressing the protein according to the expression system to obtain the protein.
According to the preparation method of the protein, the SUMO solubilizing expression label is used, and the SUMO solubilizing expression label is fused with a prokaryotic expression vector for protein expression, so that correct folding of the target protein can be better promoted, and the soluble expression quantity of the foreign protein is improved. The SUMO solubilizing expression tag is derived from a host pig, when the SUMO solubilizing expression tag is applied to preparation of the subunit vaccine for the pig, side reactions after immunization can not be caused, enzyme digestion of the SUMO solubilizing expression tag is not needed, the solubility and the expression quantity of a protective antigen can be improved, the preparation process of the subunit vaccine for the pig can be simplified, the production cost is reduced, and the production efficiency is improved.
Preferably, the protein comprises a swine disease protective antigen. Therefore, the SUMO solubilizing-promoting expression protein and the swine disease protective antigen are subjected to fusion expression in escherichia coli, a swine-derived SUMO solubilizing-promoting expression label is not required to be removed after the fusion protein is expressed, a subunit vaccine for swine can be directly prepared, after a swine is immunized, side reactions caused by the SUMO solubilizing-promoting expression label are avoided, the soluble expression of target protein can be promoted, and the effect of simplifying the preparation process of the subunit vaccine for swine can be achieved.
Specifically, the swine disease protective antigen comprises swine Japanese encephalitis virus E protein (JEV-E), swine transmissible gastroenteritis virus N protein (TGEV-N) and porcine circovirus type 3cap protein (PCV3 cap).
The invention further provides an application of the SUMO solubilizing expression label in preparing an antigen preparation. When the SUMO dissolution promoting expression tag is used for preparing an antigen preparation, the solubility and the expression quantity of antigen protein can be improved, the preparation process of the subunit vaccine for pigs can be simplified, the production cost is reduced, and the production efficiency is improved.
The technical solutions of the present invention are further described in detail below with reference to specific examples and drawings, it should be understood that the following examples are merely illustrative of the present invention and are not intended to limit the present invention.
Example 1 construction of pET28a-SUMO recombinant expression vector
(1) Analyzing and optimizing codons according to a swine-origin SUMO gene published in Genbank by using software, synthesizing an optimized SUMO sequence, namely a SUMO dissolution promotion expression label, performing PCR amplification by using specific primers (an upstream primer: 5'-cgcggatccatggcggatgaa aaaccga-3' and a downstream primer: 5'-ccgctcgagatacacgccgccggtct-3') by using the SUMO dissolution promotion expression label as a template to obtain the SUMO gene, performing gel recovery on the SUMO gene, and obtaining a agarose gel electrophoresis image, wherein M, DNAmaker2 Kprels II; lane 1, the resulting SUMO gene was PCR amplified and BamH, XhoI restriction enzyme sites were introduced at both ends.
The PCR reaction was a 50. mu.L system: mu.L of 5 XPrimeRSAR Buffer (Mg2+ Plus), 4. mu.L of dNTPmix (2.5mM), 1. mu.L of each of primers F and R (10. mu.M), 0.5. mu.L of high fidelity DNA Polymerase PrimeSTAR HS DNA Polymerase (2.5U/. mu.l), ddH2O33. mu.L, 0.5. mu.L of template diluted 10-fold. The PCR reaction conditions are as follows: 2 minutes at 98 ℃; 35 cycles of 98 ℃ for 10 seconds, 55 ℃ for 15 seconds, and 68 ℃ for 20 seconds; 10 minutes at 68 ℃. And carrying out double-enzyme digestion on the PCR recycled products of BamH and XhoI, carrying out electrophoresis analysis on the digested products by 1.5% agarose gel, and carrying out gel recovery on the PCR fragments subjected to enzyme digestion.
(2) The pET28a vector was double digested: plasmid DNA of pET28a vector was digested simultaneously with restriction nucleic acids Bam H and XhoI, the digested product was analyzed by 1.5% agarose gel electrophoresis, and vector backbone DNA fragments were recovered from the gel.
(3) And (3) connecting and transforming enzyme digestion products: the double digested SUMO gene and pET28a vector were ligated overnight at 16 ℃ using T4DNA ligase to construct expression vector pET28 a-SUMO. The ligation product was transformed into E.coli competent cells DH 5. alpha. and transformants were selected with kanamycin resistance.
(4) Verification of expression vector pET28 a-SUMO: inoculating and culturing the screened positive clone, and extracting a plasmid to perform double enzyme digestion identification of BamHI and XhoI. The recombinant clone plasmid which is correctly identified is further sequenced and identified, and the pET28a-SUMO recombinant expression vector which is correctly sequenced is stored at the temperature of 20 ℃ for later use.
Example 2 protein expression of different porcine viral protective antigens Using pET28a-SUMO recombinant expression vector
(1) pET28a and pET28a-SUMO recombinant expression vectors containing different target genes are constructed:
selecting pig Japanese encephalitis virus E protein (JEV-E), pig transmissible gastroenteritis virus N protein (TGEV-N) and pig circovirus type 3cap protein (PCV3 cap) as target proteins. Respectively amplifying target genes by PCR, introducing NheI and BamH restriction enzyme sites at two ends of PCV3cap, and introducing NdeI and BamH restriction enzyme sites at two ends of JEV-E and TGEV-N. Respectively double-digesting pET28a, pET28a-SUMO vector and PCV3cap PCR products by NheI and BamH, respectively double-digesting pET28a, pET28a-SUMO vector, JEV-E and TGEV-N PCR products by NdeI and BamH, carrying out agarose gel electrophoresis analysis on the digested products, and recovering and purifying the digested products. The double-digested target gene fragment and pET28a and pET28a-SUMO vector backbone fragments are connected at 16 ℃ overnight by using T4DNA ligase to construct recombinant expression vectors pET28a-PCV 3cap, pET28a-PCV 3cap-SUMO, pET28a-JEV-E, pET28a-JEV-E-SUMO, pET28a-TGEV-N and pET28 a-TGEV-N-SUMO. The ligation product was transformed into E.coli competent cells DH5 α and transformants were selected with kanamycin resistance. Inoculating and culturing the screened positive clone, extracting plasmid, and performing double enzyme digestion and sequencing identification. The correctly identified recombinant plasmid was stored at-20 ℃ for future use.
(2) Expression experiment of target protein:
transferring the recombinant plasmid with correct sequencing into BL21(DE3) competent cells, picking a single colony to be expressed, inoculating the single colony into a fresh Luria-Bertani liquid culture medium containing 50 mu g/ml kanamycin, culturing at 37 ℃ for 180r/min overnight, transferring the single colony to the next generation according to a ratio of 1:100 the next day, culturing at 37 ℃ until OD600 is 0.8-1.0, adding an IPTG solution with a final concentration of 0.2mM, and inducing for 17 hours at 16 ℃. The cells after induction expression were collected by centrifugation, and the cells per 100ml of the original culture were resuspended in 10ml of PBS (pH 7.4), sonicated at 0 ℃ until the cells became clear, and centrifuged at 10000rpm for 10min to collect the cells. The supernatants after centrifugation were each analyzed for differences in the soluble expression level of the target protein by 15% SDS-PAGE.
SDS-PAGE electrophoretograms of the target protein, the corresponding fusion protein and the enzyme-cleaved fusion protein are shown in FIGS. 2, 3 and 4, wherein FIG. 2 is the SDS-PAGE electrophoretogram of the JEV-E, JEV-E-SUMO fusion protein and the enzyme-cleaved JEV-E-SUMO fusion protein. In FIG. 2, M, Page Ruler Prestated Protein Ladder; lane 1, BL21(DE3) -E-SUMO E.coli induced expression after centrifugation of the supernatant; lane 2, supernatant from E.coli induced expression of BL21(DE3) -JEV-E by centrifugation; lane 3, supernatant from E.coli induced expression of enzyme BL21(DE3) -JEV-E-SUMO was centrifuged.
FIG. 3 is a SDS-PAGE electrophoresis of TGEV-N, TGEV-N-SUMO fusion protein and enzyme-cleaved TGEV-N-SUMO fusion protein. In FIG. 3, M, Page Ruler Prestated Protein Ladder; lane 2, digestion of BL21(DE3) -TGEV-N-SUMO E.coli induced expression, centrifugation of the supernatant; lane 3, BL21(DE3) -TGEV-N E.coli induced expression after centrifugation of the supernatant; lane 4, BL21(DE3) -TGEV-N-SUMO E.coli induced expression after centrifugation of the supernatant.
FIG. 4 is an SDS-PAGE electrophoresis chart of PCV3cap, PCV3cap-SUMO fusion protein and enzyme-digested PCV3cap-SUMO fusion protein. In FIG. 4, M, Page Ruler Prestated Protein Ladder; lane 1, BL21(DE3) -PCV 3cap-SUMO E.coli induced expression after centrifugation of the supernatant; lane 2, BL21(DE3) -PCV 3cap Enterobacter sp after induced expression supernatant centrifugation; lane 3, supernatant after E.coli-induced expression of enzyme BL21(DE3) -PCV 3cap-SUMO was centrifuged.
The result shows that the target protein can generate a relatively large amount of soluble protein in the genetic engineering bacteria containing the SUMO label, which indicates that the addition of the SUMO further promotes the correct folding of the expressed foreign protein and enhances the soluble expression ratio of the target protein.
In conclusion, the SUMO solubilizing expression tag provided by the invention is derived from a host pig, when the SUMO solubilizing expression tag is applied to preparation of a subunit vaccine for pigs, side reactions after immunization can not be caused, enzyme digestion of the SUMO solubilizing expression tag is not needed, the solubility and the expression quantity of a protective antigen can be improved, the preparation process of the subunit vaccine for pigs can be simplified, the production cost is reduced, and the production efficiency is improved.
The above is only a preferred embodiment of the present invention, and it is not intended to limit the scope of the invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present invention shall be included in the scope of the present invention.
SEQUENCE LISTING
<110> national drug group animal health Provisions Ltd
<120> SUMO (soluble protein synthase) promotion expression tag and application thereof, SUMO promotion expression protein, recombinant strain and preparation method of protein
Method of
<130> 2021.11.26
<160> 2
<170> PatentIn version 3.5
<210> 1
<211> 285
<212> DNA
<213> Artificial Synthesis
<400> 1
atggcggatg aaaaaccgaa agaaggcgtg aaaaccgaaa acaacgatca tattaacctg 60
aaagtggcgg gccaggatgg tagcgtggtg cagtttaaaa ttaaacgcca taccccgctg 120
agcaaactga tgaaagccta ctgcgaacgc cagggcctga gcatgcgtca gattcgcttc 180
cgtttcgatg gccagccgat taacgaaacc gataccccgg cccagctgga aatggaagat 240
gaagatacca ttgatgtgtt ccagcagcag accggcggcg tgtat 285
<210> 2
<211> 95
<212> PRT
<213> Artificial Synthesis
<400> 2
Met Ala Asp Glu Lys Pro Lys Glu Gly Val Lys Thr Glu Asn Asn Asp
1 5 10 15
His Ile Asn Leu Lys Val Ala Gly Gln Asp Gly Ser Val Val Gln Phe
20 25 30
Lys Ile Lys Arg His Thr Pro Leu Ser Lys Leu Met Lys Ala Tyr Cys
35 40 45
Glu Arg Gln Gly Leu Ser Met Arg Gln Ile Arg Phe Arg Phe Asp Gly
50 55 60
Gln Pro Ile Asn Glu Thr Asp Thr Pro Ala Gln Leu Glu Met Glu Asp
65 70 75 80
Glu Asp Thr Ile Asp Val Phe Gln Gln Gln Thr Gly Gly Val Tyr
85 90 95
Claims (8)
1. The SUMO-facilitated expression tag is characterized in that the nucleotide sequence of the SUMO-facilitated expression tag is shown in SEQ ID NO: 1 is shown.
2. The SUMO expression protein is characterized in that the amino acid sequence of the SUMO expression protein is shown as SEQ ID NO: 2, respectively.
3. A recombinant strain comprising the SUMO lysotropic expression tag of claim 1.
4. The recombinant strain of claim 3, wherein the recombinant strain comprises E.
5. A method for preparing a protein, comprising the steps of:
performing PCR amplification by using the SUMO solubilizing expression label as a template to obtain an SUMO gene;
inserting the SUMO gene into a prokaryotic expression vector through enzyme digestion reaction to obtain an expression system;
expressing the protein according to the expression system to obtain the protein;
wherein, the nucleotide sequence of the SUMO solubilizing and expressing label is shown as SEQ ID NO: 1 is shown.
6. The method of claim 5, wherein the protein comprises a swine disease protective antigen.
7. The method of claim 6, wherein the swine disease protective antigen comprises porcine encephalitis B virus E protein, porcine transmissible gastroenteritis virus N protein, and porcine circovirus type 3cap protein.
8. Use of the SUMO-facilitated expression tag of claim 1 in the preparation of an antigen formulation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111446975.3A CN114292853A (en) | 2021-11-29 | 2021-11-29 | SUMO (small cell-associated protein) lysis-promoting expression tag and application thereof, SUMO lysis-promoting expression protein, recombinant strain and preparation method of protein |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111446975.3A CN114292853A (en) | 2021-11-29 | 2021-11-29 | SUMO (small cell-associated protein) lysis-promoting expression tag and application thereof, SUMO lysis-promoting expression protein, recombinant strain and preparation method of protein |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114292853A true CN114292853A (en) | 2022-04-08 |
Family
ID=80966125
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111446975.3A Pending CN114292853A (en) | 2021-11-29 | 2021-11-29 | SUMO (small cell-associated protein) lysis-promoting expression tag and application thereof, SUMO lysis-promoting expression protein, recombinant strain and preparation method of protein |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114292853A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116731945A (en) * | 2023-07-10 | 2023-09-12 | 苏州优信合生技术有限公司 | Recombinant escherichia coli for expressing tyrosine ammonia lyase SeSam8 and application thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101679989A (en) * | 2006-12-29 | 2010-03-24 | 生命感应器公司 | Strengthen the method and composition of protein expression and purifying |
| CN108611359A (en) * | 2018-05-04 | 2018-10-02 | 武汉科前生物股份有限公司 | The preparation method and applications of 3 virus-like particle of pig circular ring virus |
| CN109593700A (en) * | 2018-12-26 | 2019-04-09 | 镇江瑞华生物科技有限公司 | A kind of method and and its application using Escherichia coli preparation bioactivity swine fever E2 albumen |
| CN109609534A (en) * | 2018-12-26 | 2019-04-12 | 镇江瑞华生物科技有限公司 | A kind of method and its application using 3 type ORF2 albumen of Bacillus coli expression pig circular ring virus |
| CN110698543A (en) * | 2019-10-18 | 2020-01-17 | 武汉科前生物股份有限公司 | Double-antigen indirect ELISA kit for African swine fever virus antibody detection |
| CN111050784A (en) * | 2017-08-11 | 2020-04-21 | 伊利诺伊大学理事会 | Truncated guinea pig L-asparaginase variants and methods of use thereof |
-
2021
- 2021-11-29 CN CN202111446975.3A patent/CN114292853A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101679989A (en) * | 2006-12-29 | 2010-03-24 | 生命感应器公司 | Strengthen the method and composition of protein expression and purifying |
| CN111050784A (en) * | 2017-08-11 | 2020-04-21 | 伊利诺伊大学理事会 | Truncated guinea pig L-asparaginase variants and methods of use thereof |
| CN108611359A (en) * | 2018-05-04 | 2018-10-02 | 武汉科前生物股份有限公司 | The preparation method and applications of 3 virus-like particle of pig circular ring virus |
| CN109593700A (en) * | 2018-12-26 | 2019-04-09 | 镇江瑞华生物科技有限公司 | A kind of method and and its application using Escherichia coli preparation bioactivity swine fever E2 albumen |
| CN109609534A (en) * | 2018-12-26 | 2019-04-12 | 镇江瑞华生物科技有限公司 | A kind of method and its application using 3 type ORF2 albumen of Bacillus coli expression pig circular ring virus |
| CN110698543A (en) * | 2019-10-18 | 2020-01-17 | 武汉科前生物股份有限公司 | Double-antigen indirect ELISA kit for African swine fever virus antibody detection |
Non-Patent Citations (9)
| Title |
|---|
| ZHONGYUAN WANG等: "Human SUMO fusion systems enhance protein expression and solubility" * |
| ZHONGYUAN WANG等: "Human SUMO fusion systems enhance protein expression and solubility", 《PROTEIN EXPRESSION AND PURIFICATION》 * |
| 李志要等: "利用猪源SUMO3标签蛋白可溶性表达猪腺病毒3型的Hexon基因高变区蛋白", 《畜牧与兽医》 * |
| 登录号：NP_008868.3: "small ubiquitin-related modifier 2 isoform a precursor [Homo sapiens]" * |
| 登录号：NP_008868.3: "small ubiquitin-related modifier 2 isoform a precursor [Homo sapiens]", 《GENBANK》 * |
| 登录号：NP_999149.1: "small ubiquitin-related modifier 2 [Sus scrofa]" * |
| 登录号：NP_999149.1: "small ubiquitin-related modifier 2 [Sus scrofa]", 《GENBANK》 * |
| 辛波等: "鸡源SUMO标签蛋白的鉴定及在IBDV VP2原核表达中的促溶功能" * |
| 辛波等: "鸡源SUMO标签蛋白的鉴定及在IBDV VP2原核表达中的促溶功能", 《中国兽医学报》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116731945A (en) * | 2023-07-10 | 2023-09-12 | 苏州优信合生技术有限公司 | Recombinant escherichia coli for expressing tyrosine ammonia lyase SeSam8 and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111217919B (en) | Novel coronavirus S protein double-region subunit nano vaccine based on pyrococcus ferritin | |
| CN111217918A (en) | Novel coronavirus S protein double-region subunit nano vaccine based on 2, 4-dioxotetrahydropteridine synthase | |
| CN112010984B (en) | Novel coronavirus S protein polymer nano vaccine based on helicobacter pylori ferritin | |
| JP2016505251A5 (en) | ||
| Ma et al. | High efficient expression, purification, and functional characterization of native human epidermal growth factor in Escherichia coli | |
| CN115838433A (en) | Beta coronavirus polymer antigen, preparation method and application thereof | |
| CN114292853A (en) | SUMO (small cell-associated protein) lysis-promoting expression tag and application thereof, SUMO lysis-promoting expression protein, recombinant strain and preparation method of protein | |
| CA2816401C (en) | Generation of antigenic virus-like particles through protein-protein linkages | |
| US20100035300A1 (en) | Producing a Target Protein Using Intramolecular Cleavage by TEV Protease | |
| KR101373297B1 (en) | Expression Vector Comprising Gene coding for E. coli Phosphoglycerate kinase As a Novel Fusion Partner | |
| KR101300672B1 (en) | Method for producing soluble foreign protein using specific intracellular cleavage system | |
| US20240093159A1 (en) | Virus-like particles and methods of production thereof | |
| CN108300725B (en) | Soluble single-chain antibody superantigen fusion gene and protein, and preparation and application thereof | |
| CN113005134B (en) | Method for promoting mass expression of glial fibrillary acidic protein in escherichia coli | |
| US10150803B2 (en) | Method of preparing glucagon-like peptide-2 (GLP-2) analog | |
| US11591630B2 (en) | Peptide for enhancing expression efficiency of target protein, and fusion protein comprising same | |
| CN104513298B (en) | A method of expression infections chicken cloacal bursa solubility VP 2-4-3 polypeptides | |
| CN114057893B (en) | Myristoylation polypeptide for encoding mitochondrial localization and preparation method and application thereof | |
| WO2019094669A2 (en) | Self-assembling protein structures and components thereof | |
| JP2023508893A (en) | Method for enhancing water solubility of target protein by WHEP domain fusion | |
| KR102120335B1 (en) | Recombinant Expression Vectors for Producing Norovirus Vaccine | |
| WO2023217286A1 (en) | Fusion protein and use thereof | |
| KR101505697B1 (en) | Membrane protein expression vector comprising major envelope protein p9 of systovirus phi12 as a fusion partner and method for producing membrane protein using the same | |
| US20230272004A1 (en) | Ramp tag for insulin overexpression and method for manufacturing insulin using same | |
| Akter et al. | Development of Recombinant Proteins for Vaccine Candidates Against Serotype O and A of Foot and Mouth Disease Virus in Bangladesh |
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 | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20220408 |
|
| RJ01 | Rejection of invention patent application after publication |