CN106636207A - Recombinant baculovirus vector resistant to host apoptosis - Google Patents
Recombinant baculovirus vector resistant to host apoptosis Download PDFInfo
- Publication number
- CN106636207A CN106636207A CN201610841372.6A CN201610841372A CN106636207A CN 106636207 A CN106636207 A CN 106636207A CN 201610841372 A CN201610841372 A CN 201610841372A CN 106636207 A CN106636207 A CN 106636207A
- Authority
- CN
- China
- Prior art keywords
- apoptosis
- host
- sequences
- caspase
- viral vector
- 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
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/86—Viral vectors
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/66—General methods for inserting a gene into a vector to form a recombinant vector using cleavage and ligation; Use of non-functional linkers or adaptors, e.g. linkers containing the sequence for a restriction endonuclease
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/14011—Baculoviridae
- C12N2710/14034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2710/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsDNA viruses
- C12N2710/00011—Details
- C12N2710/14011—Baculoviridae
- C12N2710/14041—Use of virus, viral particle or viral elements as a vector
- C12N2710/14043—Use of virus, viral particle or viral elements as a vector viral genome or elements thereof as genetic vectore
Abstract
The invention discloses a recombinant baculovirus vector resistant to host apoptosis, which is an antiapoptosis baculovirus expression vector realized by expressing siRNA of an insect cell caspase-1 through the vector. The baculovirus vector contains a section of special DNA sequence, the section of DNA sequence comprises a U6 promoter and an siRNA sequence of caspase-1 targeted by downstream 21nt thereof. A recombinant virus prepared by utilizing the vector can express double-strand small RNA in a host cell, caspase-1 coded by the host cell is silenced in an RNA interference way, and the apoptosis of the host cell can be further inhibited. The recombinant baculovirus vector can be used for constructing a recombinant baculovirus expressing an exogenous gene; after an insect cell is infected with the obtained recombinant baculovirus, the host apoptosis can be inhibited to remarkably enhance the heterologous protein expression level, thereby reducing the production cost.
Description
Technical field
The invention belongs to viral vector technology field, more particularly to a kind of recombination bacillary viral vector of anti-host's apoptosis.
Background technology
Baculoviruss are the arthropodan DNA viruses of specific infection, autographa california multiple nuclear polyhedrosis virus
(AcMNPV) be baculoviruss a type sepecies.Since nineteen eighty-three Smith etc. is transformed into expression system baculoviruss first
Since system, as the system has the advantages that low cost, high yield, and the protein conformation expressed is close to natural eukaryotic protein,
Baculovirus expression system is widely used in research and production.
Baculovirus expression system is a kind of transient expression system, and viral infected insect cell can cause cell after 3-4 days
Apoptosis and death.It is that 22-24 hours are opened from after infection based on the Time limit of expression of the baculovirus expression system of late promoter
Begin dead to host cell.If the death time of infection cell can be postponed, certainly will can increase the yield of foreign protein.
Apoptosis are that host resists one of strategy of virus infection, and wherein cell regulate and control caspase activity on cell withers
Die extremely important.And the caspase of current most study is to hold in Spodopterafrugiperda (Spodoptera frugiperda) cell
Row caspase is Sf-caspase-1.2007, a seminar in Taiwan was expressed in Sf9 cells with rna interference vector
The dsRNA of Sf-caspase-1, the Sf-caspase-1 in successful silence cell, and it is apoptotic to filter out suppression Sf9
Cell line (Lin C C, Hsu J T A, Huang K L, et al.Sf-Caspase-1-repressed stable cells:
resistance to apoptosis and augmentation of recombinant protein production
[J].Biotechnology and applied biochemistry,2007,48(1):11-19.)。
Because apoptosis is the active approaches of organism cleaning decline cell.If the anti-apoptotic approach of insect cell line itself
It is blocked, cell line will certainly be made to degenerate.The anti-apoptotic effect of current anti-apoptotic cell line does not have obvious dosage effect,
Anti-apoptotic cell line yield increase rate is less than 2 times.After current recombinate shape virus infection insect cell, insect cell pathological changes speed
Degree and degree can not be reduced, and protein expression level can not be significantly improved.
The content of the invention
It is an object of the invention to provide a kind of recombination bacillary viral vector of anti-host's apoptosis, it is intended to solve background technology
The problem for referring to.
The present invention is achieved in that a kind of recombination bacillary viral vector of anti-host's apoptosis, the weight of anti-host's apoptosis
Group baculovirus vector contains one section of specific DNA sequence, and the DNA sequence is SEQ ID NO：1.
Further, the DNA sequence includes the homologous 21nt nucleotide sequences of U6 promoteres and caspase-1,9nt loop sequences
Row, 21nt antisense sequences corresponding with 21nt nucleotide sequences, as T (n) sequences of transcription stop signalses；
Described is SEQ ID NO with caspase-1 homologous 21nt nucleotide sequences：2；
The 9nt loop sequences are SEQ ID NO：3；
The 21nt antisense sequences corresponding with 21nt nucleotide sequences are SEQ ID NO：4；
T (n) sequences as transcription stop signalses are SEQ ID NO：5.
Further, the regulating and controlling sequence that the U6 promoteres are expressed as siRNA, the U6 promoter sequences are SEQ ID NO：
6。
Another object of the present invention is to provide a kind of construction method method of the recombination bacillary viral vector of anti-host's apoptosis to be：
The coded sequence of targeting Caspase-1 tiny RNAs is directly inserted in Baculovirus Gene group；Specifically include：
1) U6 promoteres, are expanded and is cloned on vector plasmid；
2) the shRNA sequences of targeting caspase-1, are designed, and shRNA fragments are obtained with the method that long primer splices；
3) shRNA fragments are cloned into into U6 promoteres downstream,；
4), with the anti-sieve section of PCR method amplified band U6-shRNA sequences；
5), by anti-sieve section and AcMNPV Bacmid cotransformations to the escherichia coli HS996 bacterial strains containing pRed/ET, screening
Obtain the Bacmid of anti-apoptotic.
Another object of the present invention is to provide egg prepared by a kind of recombination bacillary viral vector of above-mentioned anti-host's apoptosis
White product.
Another object of the present invention is to provide epidemic disease prepared by a kind of recombination bacillary viral vector of above-mentioned anti-host's apoptosis
Seedling.
Anti-apoptotic Bacmid that the present invention is obtained, can be as viral vector, for the restructuring bar of construction expression exogenous gene
Shape virus.Because the interference RNA sequence for suppressing insect cell apoptosis, the recombinant baculovirus sense for being obtained are carried on viral vector
After dye insect cell, insect cell pathological changes speed and degree are reduced, and protein expression level is significantly improved.Jing is tested, with luciferin
Enzyme improves 10 times (see figure than the carrier luciferase activity before transformation as reporter gene, the anti-apoptotic carrier of the present invention
3).The baculovirus vector that the present invention is obtained can be used for basic research, it can also be used to the industry of large-scale protein medicine and vaccine
Metaplasia is produced.
The siRNA sequence and regulating and controlling sequence that the present invention expresses interference insect cell caspase-1 is cloned into baculoviruss
On carrier framework, the rhabdovirus expression vector that can extend infection cell survival period has been produced.
The present invention can express double-strand tiny RNA in host cell using the recombinant viruses that the carrier makes, dry by RNA
The caspase-1 of approach silence host cell coding is disturbed, so as to suppress the apoptosis of host cell.The baculovirus expression of the present invention
Carrier, can be used for the recombinant baculovirus of construction expression exogenous gene.After the recombinate shape virus infection insect cell for being obtained,
Apoptosis of Host Cells can be suppressed, significantly improve exogenous protein expression level, so as to reduce production cost.
Because apoptosis is the active approaches of organism cleaning decline cell.If the anti-apoptotic approach of insect cell line itself
It is blocked, cell line will certainly be made to degenerate.The present invention makes baculovirus vector itself have anti-apoptotic characteristic, and insect cell only has
The apoptosis pathway of cell is blocked after recombinant viruses have been infected, so as to avoid the degeneration of cell line.In addition, being different from front
Anti-apoptotic cell line is stated, the anti-apoptotic effect of the present invention has obvious dosage effect, both the baculoviruss of each cell infection
More, the tiny RNA of transcription is more, and anti-apoptotic effect is better.Based on this reason, can with the vector expression foreign protein of the present invention
So that 10 times or so of output increased, and aforementioned anti-apoptotic cell line output increased is less than 2 times.Albumen of the present invention in eukaryotic source
There is good application prospect in expression, can be used for the production of protein product and vaccine.
Description of the drawings
The construction method schematic diagram of the recombination bacillary viral vector that Fig. 1 present invention is provided.
The Western blot inspections of the expression luciferase that Fig. 2 is prepared using the recombination bacillary viral vector that the present invention is provided
Survey result.
The Enzyme activity assay result of the expression luciferase that Fig. 3 is prepared using the recombination bacillary viral vector that the present invention is provided.
Specific embodiment
In order that the objects, technical solutions and advantages of the present invention become more apparent, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that specific embodiment described herein is not used to only to explain the present invention
Limit the present invention.
Below in conjunction with the accompanying drawings the application principle of the present invention is explained in detail.
The recombination bacillary viral vector of anti-host's apoptosis provided in an embodiment of the present invention, the restructuring of anti-host's apoptosis are shaft-like
Viral vector contains one section of specific DNA sequence, and the DNA sequence is SEQ ID NO：1.
GAGGGCCTATTTCCCATGATTCCTTCATATTTGCATATACGATACAAGGCTGTTAGAGAGATAATTAGAATTAATTT
GACTGTAAACACAAAGATATTAGTACAAAATACGTGACGTAGAAAGTAATAATTTCTTGGGTAGTTTGCAGTTTTAA
AATTATGTTTTAAAATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCGATTTCTTGGCTTTATATATCTTG
TGGAAAGGACGAAGTACTGCAGGCTAGGATGCCAGTTGATAGAGCTTATTAATCTATCAACTGGCATCCTAGCTTTT
TT。
Further, the DNA sequence includes the homologous 21nt nucleotide sequences of U6 promoteres and caspase-1,9nt loop sequences
Row, 21nt antisense sequences corresponding with 21nt nucleotide sequences, as T (n) sequences of transcription stop signalses；
Described is SEQ ID NO with caspase-1 homologous 21nt nucleotide sequences：2；
GCTAGGATGCCAGTTGATAGA。
The 9nt loop sequences are SEQ ID NO：3；
GCTTATTAA。
The 21nt antisense sequences corresponding with 21nt nucleotide sequences are SEQ ID NO：4；
TCTATCAACTGGCATCCTAGC。
T (n) sequences as transcription stop signalses are SEQ ID NO：5；
TTTTTT。
Further, the regulating and controlling sequence that the U6 promoteres are expressed as siRNA, the U6 promoter sequences are SEQ ID NO：
6；
GAGGGCCTATTTCCCATGATTCCTTCATATTTGCATATACGATACAAGGCTGTTAGAGAGATAATTAGAATTAATTT
GACTGTAAACACAAAGATATTAGTACAAAATACGTGACGTAGAAAGTAATAATTTCTTGGGTAGTTTGCAGTTTTAA
AATTATGTTTTAAAATGGACTATCATATGCTTACCGTAACTTGAAAGTATTTCGATTTCTTGGCTTTATATATCTTG
TGGAAAGGACGAAGTACTGCAG。
As shown in Figure 1：The construction method of the recombination bacillary viral vector of anti-host's apoptosis provided in an embodiment of the present invention is：
The coded sequence of targeting Caspase-1 tiny RNAs is directly inserted in Baculovirus Gene group；Specifically include：
1) U6 promoteres, are expanded and is cloned on vector plasmid；
2) the shRNA sequences of targeting caspase-1, are designed, and shRNA fragments are obtained with the method that long primer splices；
3) shRNA fragments are cloned into into U6 promoteres downstream,；
4), with the anti-sieve section of PCR method amplified band U6-shRNA sequences；
5), by anti-sieve section and AcMNPV Bacmid cotransformations to the escherichia coli HS996 bacterial strains containing pRed/ET, screening
Obtain the Bacmid of anti-apoptotic.
The application principle of the present invention is further illustrated with reference to embodiment.
(1) express the apoptosis situation detection of the recombinant virus-infected cell of shRNA
The recombinant baculovirus of expression shRNA dye the method with flow cytometry with PI after infection Sf9 cells 3 days
Detection apoptosis degree.The fluorescence intensity of the cell of the recombinant virus infection of expression shRNA is substantially than comparison virus infection
Cell is weak, illustrates that the expression of shRNA can suppress the apoptotic generation that baculovirus infection causes.
(2) the Western blot detections of luciferase expression
As shown in Figure 2, after with above-mentioned viral infected insect cell 4 days, with the method detection fluorescent of Western blot
The expression of plain enzyme.As a result show, the infection for expressing the recombinant viruses of shRNA all raises the expression of luciferase,
Illustrate that inhibited apoptosis can improve protein expression level by extending the exogenous protein expression time.
(3) Enzyme activity assay of luciferase expression
As shown in Figure 3：After with above-mentioned viral infected insect cell 2-5 days, luciferase substrate detection by quantitative firefly is directly used
The expression of light element enzyme.As a result show, the infection for expressing the recombinant viruses of shRNA all makes the expression liter of luciferase
Height, after infection the 4th day expression can reach 10 times of control.
Presently preferred embodiments of the present invention is the foregoing is only, not to limit the present invention, all essences in the present invention
Any modification, equivalent and improvement made within god and principle etc., should be included within the scope of the present invention.
Claims (6)
1. a kind of recombination bacillary viral vector of anti-host's apoptosis, it is characterised in that the recombinant baculovirus of anti-host's apoptosis
Carrier contains one section of specific DNA sequence, and the DNA sequence is SEQ ID NO：1.
2. the recombination bacillary viral vector of anti-host's apoptosis as claimed in claim 1, it is characterised in that the DNA sequence includes
The U6 promoteres 21nt nucleotide sequence homologous with caspase-1,9nt loop sequences 21nt corresponding with 21nt nucleotide sequences
Antisense sequences, as T (n) sequences of transcription stop signalses；
Described is SEQ ID NO with caspase-1 homologous 21nt nucleotide sequences：2；
The 9nt loop sequences are SEQ ID NO：3；
The 21nt antisense sequences corresponding with 21nt nucleotide sequences are SEQ ID NO：4；
T (n) sequences as transcription stop signalses are SEQ ID NO：5.
3. the recombination bacillary viral vector of anti-host's apoptosis as claimed in claim 2, it is characterised in that the U6 promoteres are made
For the regulating and controlling sequence of siRNA expression, the U6 promoter sequences are SEQ ID NO：6.
4. the construction method of the recombination bacillary viral vector of the anti-host's apoptosis described in a kind of claim 1, it is characterised in that should
The construction method of the recombination bacillary viral vector of anti-host's apoptosis is：The coded sequence of targeting Caspase-1 tiny RNAs is directly inserted
Enter in Baculovirus Gene group；Specifically include：
1) U6 promoteres, are expanded and is cloned on vector plasmid；
2) the shRNA sequences of targeting caspase-1, are designed, and shRNA fragments are obtained with the method that long primer splices；
3) shRNA fragments are cloned into into U6 promoteres downstream,；
4), with the anti-sieve section of PCR method amplified band U6-shRNA sequences；
5), by anti-sieve section and AcMNPV Bacmid cotransformations to the escherichia coli HS996 bacterial strains containing pRed/ET, screening acquisition
The Bacmid of anti-apoptotic.
5. prepared by a kind of recombination bacillary viral vector of the anti-host's apoptosis using as described in claims 1 to 3 any one
Protein product.
6. prepared by a kind of recombination bacillary viral vector of the anti-host's apoptosis using as described in claims 1 to 3 any one
Vaccine.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610841372.6A CN106636207A (en) | 2016-09-22 | 2016-09-22 | Recombinant baculovirus vector resistant to host apoptosis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610841372.6A CN106636207A (en) | 2016-09-22 | 2016-09-22 | Recombinant baculovirus vector resistant to host apoptosis |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106636207A true CN106636207A (en) | 2017-05-10 |
Family
ID=58852386
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610841372.6A Pending CN106636207A (en) | 2016-09-22 | 2016-09-22 | Recombinant baculovirus vector resistant to host apoptosis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106636207A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107304433A (en) * | 2016-04-23 | 2017-10-31 | 北京康宝利华生物科技有限公司 | A kind of recombination bacillary viral vector of anti-apoptotic and preparation method thereof |
| CN108728491A (en) * | 2018-06-21 | 2018-11-02 | 陕西杆粒生物科技有限公司 | A kind of wide spectrum anti-apoptotic rhabdovirus expression vector |
| CN110387385A (en) * | 2019-07-18 | 2019-10-29 | 西北农林科技大学 | A kind of novel rhabdovirus expression vector |
| CN111378687A (en) * | 2018-12-27 | 2020-07-07 | 陕西杆粒生物科技有限公司 | High-yield baculovirus expression vector |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101260411A (en) * | 2007-12-17 | 2008-09-10 | 华中农业大学 | Recombination bacillary viral vector skeleton plasmid and application thereof |
| CN101285072A (en) * | 2008-06-05 | 2008-10-15 | 复旦大学 | Process for enhancing gene level of expression of baculoviral expression carrier |
| CN107304433A (en) * | 2016-04-23 | 2017-10-31 | 北京康宝利华生物科技有限公司 | A kind of recombination bacillary viral vector of anti-apoptotic and preparation method thereof |
-
2016
- 2016-09-22 CN CN201610841372.6A patent/CN106636207A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101260411A (en) * | 2007-12-17 | 2008-09-10 | 华中农业大学 | Recombination bacillary viral vector skeleton plasmid and application thereof |
| CN101285072A (en) * | 2008-06-05 | 2008-10-15 | 复旦大学 | Process for enhancing gene level of expression of baculoviral expression carrier |
| CN107304433A (en) * | 2016-04-23 | 2017-10-31 | 北京康宝利华生物科技有限公司 | A kind of recombination bacillary viral vector of anti-apoptotic and preparation method thereof |
Non-Patent Citations (3)
| Title |
|---|
| CHIH‐CHIEN LIN 等: "Sf-Caspase-1-repressed stable cells_ resistance to apoptosis and augmentation of recombinant protein production", 《BIOTECHNOL APPL BIOCHEM》 * |
| 欧艳梅 等: "重组杆状病毒表达Sf-caspase-1双链RNA对昆虫细胞凋亡的影响", 《西北农林科技大学(自然科学版)》 * |
| 欧艳梅: "抗宿主细胞凋亡策略以优化杆状病毒表达载体", 《中国优秀硕士学位论文全文数据库 基础科学辑》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107304433A (en) * | 2016-04-23 | 2017-10-31 | 北京康宝利华生物科技有限公司 | A kind of recombination bacillary viral vector of anti-apoptotic and preparation method thereof |
| CN108728491A (en) * | 2018-06-21 | 2018-11-02 | 陕西杆粒生物科技有限公司 | A kind of wide spectrum anti-apoptotic rhabdovirus expression vector |
| CN111378687A (en) * | 2018-12-27 | 2020-07-07 | 陕西杆粒生物科技有限公司 | High-yield baculovirus expression vector |
| CN111378687B (en) * | 2018-12-27 | 2023-06-23 | 陕西杆粒生物科技有限公司 | High-yield baculovirus expression vector |
| CN110387385A (en) * | 2019-07-18 | 2019-10-29 | 西北农林科技大学 | A kind of novel rhabdovirus expression vector |
| CN110387385B (en) * | 2019-07-18 | 2022-07-15 | 西北农林科技大学 | Baculovirus expression vector |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Cui et al. | A fungal pathogen deploys a small silencing RNA that attenuates mosquito immunity and facilitates infection | |
| CN106636207A (en) | Recombinant baculovirus vector resistant to host apoptosis | |
| Taning et al. | Engineered flock house virus for targeted gene suppression through RNAi in fruit flies (Drosophila melanogaster) in vitro and in vivo | |
| Clem et al. | Baculoviruses: sophisticated pathogens of insects | |
| Wilkins et al. | RNA interference is an antiviral defence mechanism in Caenorhabditis elegans | |
| Mascia et al. | Gene silencing and gene expression in phytopathogenic fungi using a plant virus vector | |
| Wang et al. | Suppression of Bm-Caspase-1 expression in BmN cells enhances recombinant protein production in a baculovirus expression vector system | |
| Asgari | MicroRNAs as regulators of insect host–pathogen interactions and immunity | |
| Zhang et al. | Development of a baculovirus vector carrying a small hairpin RNA for suppression of sf-caspase-1 expression and improvement of recombinant protein production | |
| Xu et al. | Knock down of target genes by RNA interference in the embryos of lepidopteran insect, Bombyx mori | |
| CN103642807B (en) | Enhanced bombyx mori nuclear polyhedrosis virus inducible promoter En39k and application thereof | |
| Flores-Jasso et al. | Silencing structural and nonstructural genes in baculovirus by RNA interference | |
| Ran et al. | Expressing microRNA bantam sponge drastically improves the insecticidal activity of baculovirus via increasing the level of ecdysteroid hormone in Spodoptera exigua larvae | |
| Kramer et al. | RNA interference as a metabolic engineering tool: potential for in vivo control of protein expression in an insect larval model | |
| CN107304433A (en) | A kind of recombination bacillary viral vector of anti-apoptotic and preparation method thereof | |
| Li et al. | Common strategies in silkworm disease resistance breeding research | |
| CN103898111B (en) | The recombinant baculovirus of the dsRNA of expression inhibiting corpus allatum hormone binding-protein gene | |
| Jia et al. | microRNA‐34‐5p encoded by Spodoptera frugiperda regulates the replication and infection of Autographa californica multiple nucleopolyhedrovirus by targeting odv‐e66, ac78 and ie2 | |
| Kim et al. | Metabolic engineering of the baculovirus‐expression system via inverse “shotgun” genomic analysis and RNA interference (dsRNA) increases product yield and cell longevity | |
| CN101285072A (en) | Process for enhancing gene level of expression of baculoviral expression carrier | |
| CN102925484A (en) | Application and recombinant vector of bombyx mori nuclear polyhedrosis virus polygene inverted repeat sequence and bombyx mori lipase-1 gene | |
| CN108728491B (en) | Broad-spectrum anti-apoptosis baculovirus expression vector | |
| Enden et al. | A model of the dynamics of insect cell infection at low multiplicity of infection | |
| CN102154290A (en) | SiRNAs for inhibiting epidemic encephalitis B viruses | |
| Singh | Viral-encoded microRNAs in host-pathogen interactions in silkworm |
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 | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20170510 |