WO2015109953A1 - 一种人源抗虫基因及其编码的抗Cry1C毒素独特型单链抗体与应用 - Google Patents

一种人源抗虫基因及其编码的抗Cry1C毒素独特型单链抗体与应用 Download PDF

Info

Publication number
WO2015109953A1
WO2015109953A1 PCT/CN2015/070422 CN2015070422W WO2015109953A1 WO 2015109953 A1 WO2015109953 A1 WO 2015109953A1 CN 2015070422 W CN2015070422 W CN 2015070422W WO 2015109953 A1 WO2015109953 A1 WO 2015109953A1
Authority
WO
WIPO (PCT)
Prior art keywords
insect
chain antibody
resistant gene
cry1c
anthropogenic
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.)
Ceased
Application number
PCT/CN2015/070422
Other languages
English (en)
French (fr)
Chinese (zh)
Inventor
刘贤金
徐重新
张霄
刘媛
谢雅晶
张存政
余向阳
王冬兰
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Jiangsu Academy of Agricultural Sciences
Original Assignee
Jiangsu Academy of Agricultural Sciences
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Jiangsu Academy of Agricultural Sciences filed Critical Jiangsu Academy of Agricultural Sciences
Priority to BR112016016027-4A priority Critical patent/BR112016016027B1/pt
Priority to US15/025,805 priority patent/US9745385B2/en
Priority to EP15740473.2A priority patent/EP3048171B1/en
Priority to JP2016535246A priority patent/JP6211703B2/ja
Publication of WO2015109953A1 publication Critical patent/WO2015109953A1/zh
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Images

Classifications

    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/50—Isolated enzymes; Isolated proteins
    • 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
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N47/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid
    • A01N47/40—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid the carbon atom having a double or triple bond to nitrogen, e.g. cyanates, cyanamides
    • A01N47/42—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom not being member of a ring and having no bond to a carbon or hydrogen atom, e.g. derivatives of carbonic acid the carbon atom having a double or triple bond to nitrogen, e.g. cyanates, cyanamides containing —N=CX2 groups, e.g. isothiourea
    • A01N47/44—Guanidine; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N57/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds
    • A01N57/10—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-oxygen bonds or phosphorus-to-sulfur bonds
    • A01N57/16—Biocides, pest repellants or attractants, or plant growth regulators containing organic phosphorus compounds having phosphorus-to-oxygen bonds or phosphorus-to-sulfur bonds containing heterocyclic radicals
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/12—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from bacteria
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/12—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from bacteria
    • C07K16/1267—Gram-positive bacteria
    • C07K16/1278—Bacillus (G)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies
    • C07K16/42—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against immunoglobulins
    • C07K16/4208—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an idiotypic determinant on Ig
    • C07K16/4233—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against immunoglobulins against an idiotypic determinant on Ig against anti-bacterial Ig
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
    • C07K2317/622—Single chain antibody (scFv)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding

Definitions

  • the invention relates to the field of genetic engineering and biological control, in particular to a human-resistant insect-resistant gene and an encoded anti-Cry1C toxin unique type single-chain antibody and application thereof.
  • Bacillus thuringiensis Bacillus thuringiensis
  • Bacillus thuringiensis Bacillus thuringiensis is an insect pathogenic bacterium, and its Bt toxin has a specific poisoning effect on a variety of agricultural and forest pests.
  • Plant Genetic Systems of Belgium reported the success of transgenic Bt insect-resistant tobacco for the first time in the world.
  • Bt gene has been transferred to corn, rice, cotton, tomato, potato, tobacco, etc. Among the world's major crops.
  • Bt toxin protein In the liver and kidney, cell swelling and vacuolar-like degeneration can be observed, and Seeing the lesions of glomerular vascular epithelial cells, of course, this cannot be ruled out by the immune response. Long-term high-dose use of Bt toxin protein also resulted in a significant decrease in the total number of white blood cells and hemoglobin in animals, indicating that Bt toxin protein has significant immunosuppressive toxicity. The development of alternative biological effectors with Bt toxin bioactivity (eg anti-idiotypic antibodies) is a hot spot in the development of biopesticide research.
  • An anti-idiotype antibody (hereinafter referred to as Anti-Id) is a specific antibody produced by a unique type (Idiotype, Id) located in the variable region of an antibody molecule.
  • Id an anti-idiotype antibody
  • Bona et al. classify anti-idiotypic antibodies into four types ( ⁇ , ⁇ , ⁇ , and ⁇ ) based on the serological response of Id and Anti-Id and the function of AId.
  • the ⁇ -type anti-idiotypic antibody has an antigenic function and biological activity because it has an "inner image" effect, that is, has the same antigenic determinant as the (semi)antigen.
  • phage display technology can be used to construct an anti-idiotypic antibody with a similar effect of a target antigen by constructing a phage antibody library.
  • the process of screening for specific antibodies using phage display technology is called “Panning” and consists of four steps of binding, washing, elution and amplification.
  • Raats et al. used anti-cortisol monoclonal anti-coating as a solid phase antigen for direct screening, and negative screening with negative monoclonal antibodies of the same species before screening to avoid screening for recombinant antibody fragments that bind to the constant region of the antibody.
  • the anti-Id against cortisol was successfully screened. Goletz et al.
  • Anti-Id ScFvs anti-Bt toxin type anti-idiotype single-chain antibodies
  • a human insect-resistant gene having a nucleotide sequence as shown in SEQ ID NO.
  • the anti-Cry1C toxin idiotype single-chain antibody encoded by SEQ ID NO. 1 has an amino acid sequence as shown in SEQ ID NO.
  • a prokaryotic expression vector comprising the human insect resistance gene SEQ ID NO.
  • a human insect resistance gene SEQ ID NO. 1 is used in the control of agricultural pests.
  • an insecticide comprising an anti-Cry1C toxin idiotype single-chain antibody having an amino acid sequence as shown in SEQ ID NO.
  • the invention screens and obtains an insecticidal activity " ⁇ " type anti-Cry1C toxin unique type single chain antibody from the published human gene library, and the single chain antibody is expressed by a prokaryotic system, and the primary culture has a vertical volume with rice
  • the binding activity of the peritrophic membrane specific receptor BBMV in the midgut of the leaf mites the invention obtains the " ⁇ " type anti-Cry1C toxin unique type single-chain antibody without animal immunization, has a short preparation cycle, small amino acid sequence, and is suitable for large scale in vitro.
  • the present invention as a brand new new insect-resistant genetic resource, explores and expands new insect-resistant genetic resources with simulated Bt toxin biological activity, reduces various safety risks existing in the widespread use of existing Bt toxins, and may replace Bt in the future. It is of great scientific and practical significance to prevent and control the use of pesticides in biological control of agricultural pests.
  • Figure 1 is a schematic diagram of the results of the E8 ELISA assay.
  • Figure 2 is a schematic diagram showing the results of biological measurement of E8.
  • Fig. 3 is a schematic diagram showing the death of the third instar larvae of Cnaphalocrocis medinalis after being fed with E8, CK+ and CK-soaked rice leaves, respectively.
  • Figure 4 shows the death of third instar larvae of Plutella xylostella after feeding E8, CK+, CK-soaked cabbage slices Schematic diagram.
  • a final concentration of 100 ⁇ g/ml ampicillin and a mass ratio of 1% glucose were added to the 2 ⁇ TY medium.
  • a final concentration of 100 ⁇ g/ml ampicillin and 50 ⁇ g/ml kanamycin was added to the 2 ⁇ TY medium.
  • a final concentration of 100 ⁇ g/ml ampicillin, 50 ⁇ g/ml kanamycin, and a mass ratio of 1% glucose were added to the 2 ⁇ TY medium.
  • a final concentration of 100 ⁇ g/ml ampicillin was added to the TYE solid medium and the mass ratio was 1%. glucose.
  • Tween-20 in a volume ratio of 0.05% was added to the PBS solution.
  • Citrate buffer (CPBS, substrate buffer, pH 5.5):
  • Anti-Cry1C polyclonal antibody, BBMV, unrelated anti-idiotype single-chain antibody, non-" ⁇ " type Anti-IdScFv, cabbage leaf, Plutella xylostella third instar larvae are all controlled by agricultural quality control technology of the Ministry of Agriculture of Jiangsu Academy of Agricultural Sciences Provided by standard focus laboratories;
  • Humanized phage antibody library, TG1 bacteria and helper phage KM13 were purchased from UK Source BioScience;
  • HRP-goat anti-M13-IgG was purchased from Wuhan Boster Company;
  • Cry1C toxin and Cry1Ab toxin were purchased from Shanghai Youlong Biotechnology Co., Ltd.;
  • the third instar larvae of rice leaves and leaf roller are provided by Yangzhou Luyuan Biochemical Co., Ltd.
  • the eluate is the phage antibody in the first round of panning; 2, 3,
  • the concentration of the anti-Cry1C polyclonal antibody in the four rounds of panning was 50 ⁇ g/ml, 25 ⁇ g/ml, and 10 ⁇ g/ml, respectively.
  • the phage antibody used was the phage antibody obtained by the previous round of panning, the panning method and the first round.
  • the cells were cultured overnight; the next day, 2 ⁇ l of the bacterial solution was aspirated from the wells and transferred to a new 96-well plate well, incubated at 37 ° C for 2 h, and 25 ⁇ l of helper phage KM13 with a titer of 10 12 was added to each well, incubated at 30 ° C for 2 h, centrifuged at 1800 g. After 10 min, the pellet was resuspended in 150 ⁇ l of 2 ⁇ TY-AK liquid medium, cultured at 30 ° C overnight, and centrifuged at 1800 g for 30 min the next day, and the supernatant was separately taken;
  • the nucleotide sequence of the selected anti-Cry1C toxin idiotype single-chain antibody was determined by Sanger sequencing method as SEQ ID NO. 1 as follows:
  • the translated amino acid sequence of the anti-Cry1C toxin idiotype single-chain antibody was determined by Sanger sequencing method as SEQ ID NO. 2, as follows:
  • the supernatant containing the anti-Cry1C toxin unique type single-chain antibody screened in Example 1 was transferred to 10 ml of 2 ⁇ TY-AG liquid medium at a volume ratio of 1:100, incubated at 37 ° C for 2 h, and a titer of 100 ⁇ l was added.
  • the experiment was divided into 6 experimental groups and corresponding control groups, and the solutions were prepared according to Table 1.
  • E8 in Table 1 is the supernatant of the E8-containing primary culture obtained in Example 2;
  • the experiment was divided into experimental group 1, experimental group 2, experimental group 3, positive control group, negative control group 1, negative control group 2, and negative control group 3; the experimental steps were as follows:
  • E8 is the supernatant of the E8-containing primary culture obtained in Example 2;
  • the experiment was divided into experimental group and control group:
  • the experimental group obtained the supernatant (E8) containing the E8 primary culture obtained in Example 2;
  • the positive control group used Cry1Ab toxin (CK+) at a concentration of 0.2 g/L;
  • the negative control group used anti-Id ScFvs (CK-) other than " ⁇ " type;
  • Figures 3 and 4 The experimental results are shown in Figures 3 and 4, wherein Figure 3 is fed with E8, Cry1Ab toxin (CK+), The death of the third instar larvae of rice leaf roller, which is not in the " ⁇ " type of Anti-Id ScFvs (CK-) soaked rice leaves, Figure 4 is the feeding of E8, Cry1Ab toxin (CK+), non-" ⁇ ", respectively. The death of the third instar larvae of Plutella xylostella after the type of Anti-Id ScFvs (CK-) soaked cabbage tablets shows that E8 has a good insecticidal effect.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Genetics & Genomics (AREA)
  • Zoology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Wood Science & Technology (AREA)
  • Immunology (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Molecular Biology (AREA)
  • Plant Pathology (AREA)
  • Medicinal Chemistry (AREA)
  • Dentistry (AREA)
  • Agronomy & Crop Science (AREA)
  • Environmental Sciences (AREA)
  • Pest Control & Pesticides (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biomedical Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Virology (AREA)
  • Physics & Mathematics (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Peptides Or Proteins (AREA)
  • Catching Or Destruction (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
PCT/CN2015/070422 2014-01-26 2015-01-09 一种人源抗虫基因及其编码的抗Cry1C毒素独特型单链抗体与应用 Ceased WO2015109953A1 (zh)

Priority Applications (4)

Application Number Priority Date Filing Date Title
BR112016016027-4A BR112016016027B1 (pt) 2014-01-26 2015-01-09 Gene de resistência a inseto derivado de humanos e anticorpo de cadeia única de idiotipo anti-toxina crylc codificado pelo mesmo e uso do mesmo
US15/025,805 US9745385B2 (en) 2014-01-26 2015-01-09 Anthropogenic insect-resistant gene and Cry1C toxin idiotype single-chain antibody encoded thereby and application thereof
EP15740473.2A EP3048171B1 (en) 2014-01-26 2015-01-09 Anthropogenic insect-resistant gene and cry1c toxin idiotype single-chain antibody encoded thereby and application thereof
JP2016535246A JP6211703B2 (ja) 2014-01-26 2015-01-09 ヒト耐虫遺伝子およびそれによりコードされた抗Ｃｒｙ１Ｃ毒素イディオタイプ一本鎖抗体ならびに用途

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201410037240.9A CN103773776B (zh) 2014-01-26 2014-01-26 一种人源抗虫基因及其编码的抗Cry1C毒素独特型单链抗体与应用
CN201410037240.9 2014-01-26

Publications (1)

Publication Number Publication Date
WO2015109953A1 true WO2015109953A1 (zh) 2015-07-30

Family

ID=50566541

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2015/070422 Ceased WO2015109953A1 (zh) 2014-01-26 2015-01-09 一种人源抗虫基因及其编码的抗Cry1C毒素独特型单链抗体与应用

Country Status (6)

Country Link
US (1) US9745385B2 (enExample)
EP (1) EP3048171B1 (enExample)
JP (1) JP6211703B2 (enExample)
CN (1) CN103773776B (enExample)
BR (1) BR112016016027B1 (enExample)
WO (1) WO2015109953A1 (enExample)

Families Citing this family (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103773775B (zh) 2014-01-26 2015-11-18 江苏省农业科学院 一种人源抗虫基因及其编码的抗Cry1B毒素独特型单链抗体与应用
CN104498501B (zh) * 2014-12-05 2017-07-18 江苏省农业科学院 一种人源杀虫基因及其编码杀虫肽与应用
CN104926940B (zh) * 2015-06-15 2018-02-09 江苏省农业科学院 一种人源杀虫蛋白及其制备方法与应用
CN105131123B (zh) * 2015-09-16 2018-11-27 江苏省农业科学院 一种具有杀虫活性的牛源化改型抗体及其制备方法与应用
CN105085683B (zh) * 2015-09-16 2018-11-27 江苏省农业科学院 一种具有杀虫活性的猪源化改型抗体及其制备方法与应用
EP3415010A1 (en) 2017-06-13 2018-12-19 Agrosavfe Nv Insect-controlling polypeptides and methods

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040221334A1 (en) * 1996-11-27 2004-11-04 Monsanto Technology Llc. Antibodies immunoreactive with lepidopteran-toxic polypeptides and methods of use
CN102936598A (zh) * 2012-11-21 2013-02-20 江苏省农业科学院 抗Cry1Ac毒素单链抗体的编码基因及免疫PCR检测方法

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US7060491B1 (en) * 1999-11-18 2006-06-13 Pioneer Hi-Bred International, Inc. Polynucleotides encoding novel BT toxin receptors from lepidopteran insects
WO2007023782A1 (ja) * 2005-08-24 2007-03-01 Genecare Research Institute Co., Ltd. キラートキシン様タンパク質に対する中和抗体、およびその利用

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040221334A1 (en) * 1996-11-27 2004-11-04 Monsanto Technology Llc. Antibodies immunoreactive with lepidopteran-toxic polypeptides and methods of use
CN102936598A (zh) * 2012-11-21 2013-02-20 江苏省农业科学院 抗Cry1Ac毒素单链抗体的编码基因及免疫PCR检测方法

Non-Patent Citations (6)

* Cited by examiner, † Cited by third party
Title
LING LIU ET AL.: "Influence of Transgenic Bt Maize on Bacterial Quantity Diversity of Soil", JOURNAL OF ECOLOGY AND RURAL ENVIRONMENT, 2011
MIN WANG ET AL.: "Diversity of Rhizospheric Microorganisms and Bacterial Physiological Groups of Transgenic Bt Maize", CHINESE JOURNAL OF ECOLOGY, 2010
See also references of EP3048171A4
VAZQUEZ-PADRON ET AL.: "CrylAc protoxin from Bacillus thuringiensis sp. kurstaki HD73 binds to surface proteins in the mouse small intestine", BIOCHEM BIOPHYS RES COMMUN, 2000
WANG, YUN ET AL.: "Isolation of single chain variable fragment (scFv) specific for CrylC toxin from human single fold scFv libraries", TOXICON., vol. 60, no. 7, 5 September 2012 (2012-09-05), pages 1290 - 1297, XP028947202 *
WANG, YUN ET AL.: "Screening of anti-CrylAc scFv from A phage display antibody library", CHINESE JOURNAL OF CELLULAR AND MOLECULAR IMMUNOLOGY, vol. 25, no. 12, 31 December 2009 (2009-12-31), XP008183116 *

Also Published As

Publication number Publication date
BR112016016027B1 (pt) 2022-02-08
EP3048171A1 (en) 2016-07-27
JP6211703B2 (ja) 2017-10-11
US9745385B2 (en) 2017-08-29
JP2017505111A (ja) 2017-02-16
US20160215066A1 (en) 2016-07-28
CN103773776B (zh) 2015-11-18
BR112016016027A2 (pt) 2017-12-12
EP3048171A4 (en) 2017-03-01
EP3048171B1 (en) 2018-05-02
CN103773776A (zh) 2014-05-07

Similar Documents

Publication Publication Date Title
WO2015109951A1 (zh) 一种人源抗虫基因及其编码的抗Cry1Ab毒素独特型单链抗体与应用
CN103773775B (zh) 一种人源抗虫基因及其编码的抗Cry1B毒素独特型单链抗体与应用
EP3048171B1 (en) Anthropogenic insect-resistant gene and cry1c toxin idiotype single-chain antibody encoded thereby and application thereof
WO2016086513A1 (zh) 一种人源杀虫基因及其编码杀虫肽与应用
CN116178512B (zh) 一种模拟Bt Cry毒素共性结构和功能的多肽及其编码基因与应用
Shen et al. Screening and identification of anti-idiotypic nanobody capable of broad-spectrum recognition of the toxin binding region of lepidopteran cadherins and mimicking domain II of Cry2Aa toxin
CN105884892B (zh) 一种Bt Cry毒素广谱检测用蛋白及其编码基因与应用
CN113912684B (zh) 一种模拟Bt Cry1Ac蛋白关键抗虫功能表位的短肽及其编码基因与应用
CN118930646B (zh) 一种具备模拟Bt Cry1C毒素结构和杀虫功能的人源基因工程抗体及其突变体
CN105085683A (zh) 一种具有杀虫活性的猪源化改型抗体及其制备方法与应用
CN121108329A (zh) 一种人源单域抗体抗虫基因c4d及其编码的蛋白质人源单域抗体c4d和应用

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 15740473

Country of ref document: EP

Kind code of ref document: A1

WWE Wipo information: entry into national phase

Ref document number: 15025805

Country of ref document: US

REEP Request for entry into the european phase

Ref document number: 2015740473

Country of ref document: EP

WWE Wipo information: entry into national phase

Ref document number: 2015740473

Country of ref document: EP

ENP Entry into the national phase

Ref document number: 2016535246

Country of ref document: JP

Kind code of ref document: A

REG Reference to national code

Ref country code: BR

Ref legal event code: B01A

Ref document number: 112016016027

Country of ref document: BR

NENP Non-entry into the national phase

Ref country code: DE

REG Reference to national code

Ref country code: BR

Ref legal event code: B01E

Ref document number: 112016016027

Country of ref document: BR

ENP Entry into the national phase

Ref document number: 112016016027

Country of ref document: BR

Kind code of ref document: A2

Effective date: 20160708