WO2021073220A1 - 具有除草剂麦草畏降解功能的基因dicX4及其应用 - Google Patents

具有除草剂麦草畏降解功能的基因dicX4及其应用 Download PDF

Info

Publication number
WO2021073220A1
WO2021073220A1 PCT/CN2020/108309 CN2020108309W WO2021073220A1 WO 2021073220 A1 WO2021073220 A1 WO 2021073220A1 CN 2020108309 W CN2020108309 W CN 2020108309W WO 2021073220 A1 WO2021073220 A1 WO 2021073220A1
Authority
WO
WIPO (PCT)
Prior art keywords
dicx4
gene
herbicide
dicamba
strain
Prior art date
Application number
PCT/CN2020/108309
Other languages
English (en)
French (fr)
Inventor
林敏�
周正富
张维
燕永亮
陈明
陆伟
Original Assignee
中国农业科学院生物技术研究所
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 中国农业科学院生物技术研究所 filed Critical 中国农业科学院生物技术研究所
Publication of WO2021073220A1 publication Critical patent/WO2021073220A1/zh

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • 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
    • C12N15/82—Vectors or expression systems specially adapted for eukaryotic hosts for plant cells, e.g. plant artificial chromosomes (PACs)
    • C12N15/8241—Phenotypically and genetically modified plants via recombinant DNA technology
    • C12N15/8261—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield
    • C12N15/8271—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance
    • C12N15/8274—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for stress resistance, e.g. heavy metal resistance for herbicide resistance

Definitions

  • the invention relates to a gene with the function of degrading the herbicide dicamba.
  • Dicamba is a selective and systemic conductive post-emergence auxin herbicide. In 1961, dicamba was created by Velsicol Chemical Corporation, with a chemical name of 3,6-dichloro-2-methoxybenzoic acid, CAS: 1918-00-9, and a molecular weight of 221.04.
  • dicamba has good systemic conductivity, it can be absorbed by the stems, leaves or roots of weeds within a short period of time, and the weeds can be killed by roots. It is stable and easy to move in the soil, and has a certain residual period, which can generally be maintained for more than 40 days, causing its residue in the soil to be very serious. At present, the degradation mechanism of dicamba is not clear, and it is impossible to study the target protein. Only part of the research was carried out on the cloning and acquisition of the dicamba degrading enzyme gene.
  • microorganisms After long-term pressure selection, microorganisms have evolved a series of survival mechanisms and functional genes that adapt to the habitat. The genomes of these strains contain rich genetic resources, which are worthy of in-depth study.
  • the purpose of the present invention is to discover a new herbicide dicamba degradation gene, the protein encoded by it can be used for the degradation of dicamba molecules.
  • the present invention obtains a gene dicX4 which can degrade the herbicide dicamba from the microbial metagenomic data of soil samples.
  • a gene dicX4 which can degrade the herbicide dicamba from the microbial metagenomic data of soil samples.
  • the gene was expressed in the herbicide degradation screening strain Escherichia coli.
  • the gene has the function of degrading herbicide dicamba after being expressed in prokaryotic host cells, and can be used for the cultivation of herbicide-resistant transgenic crops and the bioremediation of contaminated soil.
  • the specific research work is as follows:
  • the dicX4 gene was amplified by PCR from the microbial metagenomics of a soil sample in southern China. Its size is 1086bp. The gene encodes 361 amino acids. It was cloned into the vector pJET to construct a recombinant clone containing the complete dicX4 gene. Plasmid pJET-dicX4;
  • the recombinant engineered strain JM-dicX4 expressing the dicX4 gene can grow in a medium with dicamba as the sole carbon source, and has the ability to degrade the herbicide dicamba. After adding the herbicide dicamba compound to the medium, after 48 hours of incubation, the recombinant E. coli engineered strain JM-dicX4 expressing dicX4 can completely degrade the dicamba molecule to produce demethylated 3,6-non-herbicidally active 3,6- Dichlorosalicylic acid (DCSA).
  • DCSA demethylated 3,6-non-herbicidally active 3,6- Dichlorosalicylic acid
  • DicX4 protein has the catalytic function of degrading the herbicide dicamba, and the degradation product is mainly DCSA without herbicide activity.
  • the gene has application potential in the cultivation of herbicide-resistant transgenic crops and the bioremediation of contaminated soil.
  • SEQ ID NO. 1 The nucleotide sequence of the dicX4 gene.
  • SEQ ID NO. 2 The amino acid sequence of DicX4.
  • Figure 1 The recombinant engineering strain JM-dicX4 can metabolize dicamba.
  • FIG. 1 HPLC detection diagram of standard dicamba and 3,6-dichlorosalicylic acid, in which:
  • Figure 3B shows that after 48 hours of incubation, the main product of the recombinant engineered strain JM-dicX4 (solid line) is DCSA (peak No. 2). However, the control strain JM-D1 (dotted line) did not detect the No. 2 peak substance, and had no dicamba degradation ability.
  • the plasmids, strains, and microorganisms to be catalyzed and degraded in the following examples are only used to further describe the present invention in detail, and do not limit the essence of the present invention. Where the specific experimental conditions are not indicated, they are in accordance with the conventional conditions well known to those skilled in the art or the conditions suggested by the manufacturer.
  • the sources of plasmids and strains cited in the examples are as follows:
  • Cloning vector pJET a commercial product of ThermoFisher
  • Escherichia coli JM109 It is a commercial product of Beijing Quanshijin Company.
  • Standard dicamba and 3,6-dichlorosalicylic acid commercial products of sigma-aldrich company.
  • Escherichia coli JM109 It is a commercial product of Beijing Quanshijin Company.
  • dicX4-F 5'ACCACTAGTATGCCTTTCGTTTACAATGC 3'
  • Reaction conditions 95°C for 10min, [95°C for 30sec, 61°C for 30sec, 72°C for 1.0min] 35 cycles, 72°C for 10min.
  • the dicX4 gene in the soil metagenomics was successfully cloned using PCR technology, and a recombinant E. coli engineered strain expressing the dicX4 gene was successfully constructed.
  • the correct insertion sequence was verified by PCR, restriction enzyme digestion, and sequencing, and the strain was named JM-dicX4.
  • E. coli JM109 containing the pRAD1 control empty plasmid was named JM-D1.
  • Recombinant engineering strain JM-dicX4 strain expressing dicX4 gene obtained in Example 1
  • Control strain the JM-D1 strain described in Example 1 containing the empty plasmid.
  • the recombinant engineered strain JM-dicX4 can metabolize dicamba.
  • the recombinant engineered strain JM-dicX4 that expresses the dicX4 gene in Figure 1 can grow in a medium with dicamba as the sole carbon source, and the medium becomes turbid. After 48 hours of incubation, the dicamba is degraded and the color of the medium gradually turns red.
  • Figure 1 The control strain JM-D1 containing the empty plasmid after 48h incubation, the medium is still clear and no color change is seen.
  • the recombinant engineered strain JM-dicX4 expressing the dicX4 gene can grow in a medium with dicamba as the sole carbon source, and has the ability to degrade the herbicide dicamba.
  • Recombinant engineering strain JM-dicX4 strain expressing dicX4 gene obtained in Example 1
  • Control strain the JM-D1 strain described in Example 1 containing the empty plasmid.
  • the chromatographic column is a C18 column.
  • the sample loading volume was 100 ⁇ L, and dicamba and its degradation products were diluted and analyzed by using the gradient change of the mobile phase.
  • the gradient of mobile phase solution B is from 30%-95% in 30min.
  • A: ultrapure water: acetonitrile: methanol: acetic acid 58.4:31.7:7.5:2.4; B: 100% acetonitrile
  • the flow rate is 0.8 mL/min.
  • the recombinant engineered strain JM-dicX4 expressing the dicX4 gene and the control strain JM-D1 strain containing an empty plasmid have significantly different product peak patterns.
  • the main compound peak in the medium of the control strain JM-D1 (dotted line) is still peak No. 1, with a retention time of 6 min.
  • the peak of the control standard product is a dicamba compound.
  • the compound peak pattern in the medium of the recombinant engineered strain JM-dicX4 (solid line) expressing the dicX4 gene has changed significantly, and peak 1 has not been detected.
  • Figure 3B shows that after 48 hours of incubation, the main product of the recombinant engineered strain JM-dicX4 (solid line) is peak No. 2 with a retention time of 5.1 min.
  • the peak of the reference standard product is a DCSA compound.
  • the control strain JM-D1 (dotted line) did not detect the production of peak 2 substance.
  • the recombinantly expressed DicX4 protein has the catalytic function of degrading the herbicide dicamba, and the degradation product is mainly 3,6-dichlorosalicylic acid (DCSA) without herbicide activity.
  • DCSA 3,6-dichlorosalicylic acid

Landscapes

  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Organic Chemistry (AREA)
  • Molecular Biology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biophysics (AREA)
  • Biotechnology (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Plant Pathology (AREA)
  • Cell Biology (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Medicinal Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

提供了一种从土壤样品的宏基因组中获得的具有降解除草剂麦草畏的基因dicX4，还构建了含有该基因的重组载体，并将其在原核宿主细胞大肠杆菌中表达。实验证明，该基因具有降解除草剂麦草畏的功能，能用于抗除草剂转基因农作物的培育及污染土壤的生物修复。

Description

具有除草剂麦草畏降解功能的基因dicX4及其应用 技术领域
本发明涉及一种具有降解除草剂麦草畏功能的基因。
背景技术
麦草畏(Dicamba)是一种具有选择性和内吸传导型苗后生长素类除草剂。1961年，麦草畏由美国维尔斯科尔化学公司(Velsicol Chemical Corporation)创制，化学名为3，6-二氯-2-甲氧基苯甲酸，CAS：1918-00-9，分子量为221.04。
由于麦草畏的内吸传导性好，在较短时间内可被杂草的茎、叶或根内吸，可以将杂草连根杀死。在土壤中稳定且较易移动，有一定的残效期，一般能保持40天以上，造成其在土壤中的残留十分严重。目前，对麦草畏的降解机制还不清晰，无法针对靶蛋白进行研究。仅针对麦草畏降解酶基因的克隆和获取开展了部分研究。
微生物经过长期的压力选择，进化出一系列适应该生境的生存机制与功能基因。这些菌株基因组中蕴藏着丰富的基因资源，值得进行深入研究。
发明内容
本发明的目的是发现一种新的除草剂麦草畏降解基因，其编码的蛋白质能够用于麦草畏分子的降解。
本发明从土壤样品的微生物宏基因组数据中获得了一种具有降解除草剂麦草畏的基因dicX4。通过构建该基因的表达载体，将其在除草剂降解筛选菌株大肠杆菌中表达。实验证明，所述基因在原核宿主细胞中表达后，具有降解除草剂麦草畏的功能，可用于抗除草剂转基因农作物的培育及污染土壤的生物修复。具体研究工作如下：
1、获得了含有dicX4基因的重组工程菌株
1)通过PCR从中国南方某地土壤样品的微生物宏基因组扩增出dicX4基因，其大小为1086bp，该基因编码361个氨基酸，将其克隆于载体pJET上，构建了含有完整dicX4基因的重组克隆质粒pJET-dicX4；
2)将dicX4基因连接于pRAD1质粒上，该质粒含组成型高表达groEL启动子，可高效持续表达下游靶标蛋白，完成重组质粒pRAD-dicX4的构建；
3)将导入dicX4基因的重组质粒pRAD-dicX4转入麦草畏降解基因筛选菌株大肠杆菌中，获得工程菌株JM-dicX4(详见实施例1)；
2、表达dicX4的重组大肠杆菌工程菌株的催化活性实验
已有研究表明，除草剂麦草畏降解的关键步骤在于麦草畏分子的去甲氧基化反应。经过微生物酶的催化将除草剂麦草畏去除甲氧基基团生成无除草剂活性的3,6-二氯水杨酸(DCSA)。
实验证实，表达dicX4基因的重组工程菌JM-dicX4能够在以麦草畏为唯一碳源的培养基中生长，具有降解除草剂麦草畏的能力。向培养基中添加除草剂麦草畏化合物后，经过48小时孵育，表达dicX4的重组大肠杆菌工程菌株JM-dicX4能够将麦草畏分子完全降解，生成去甲基的无除草剂活性的3,6-二氯水杨酸(DCSA)。
实验表明，重组表达的DicX4蛋白质具有降解除草剂麦草畏的催化功能，降解产物主要为无除草剂活性的DCSA。该基因在抗除草剂转基因农作物的培育及污染土壤的生物修复方面具有应用潜力。
序列表信息
SEQ ID NO.1：dicX4基因的核苷酸序列。
SEQ ID NO.2：DicX4的氨基酸序列。
附图说明：
图1重组工程菌JM-dicX4能够代谢麦草畏。
图2标准品麦草畏与3,6-二氯水杨酸的HPLC检测图，其中：
A，麦草畏，检测波长λ＝275nm，出峰时间6min；
B，DCSA，检测波长为λ＝319nm，出峰时间为5.1min；
图3工程菌株JM-dicX4对麦草畏降解效果：
图3A中，48h孵育培养后，表达dicX4基因的重组工程菌JM-dicX4(实线)培养基中麦草畏(1号峰)已经检测不到；
图3B中显示，48h孵育培养后，重组工程菌JM-dicX4(实线)的主要产物为DCSA(2号峰)。而对照菌株JM-D1(虚线)未检测到2号峰物质，无麦草畏降解能力。
具体实施方式
以下实施例中所举的质粒、菌株以及微生物催化降解的对象只用于对本发明作进一步详细说明，并不对本发明的实质内容加以限制。凡未注明具体实验条件的，均为按照本领域技术人员熟知的常规条件或按照制造厂商所建议的条件。实施例中所举的质粒、菌株来源如下：
克隆载体pJET：为ThermoFisher公司市售产品；
表达质粒pRAD1：本实验室保藏；
大肠杆菌JM109：为北京全式金公司市售产品。
标准品麦草畏与3,6-二氯水杨酸：为sigma-aldrich公司市售产品。
实施例1 土壤宏基因组中dicX4基因序列在大肠杆菌中的表达
一、实验材料
大肠杆菌JM109：为北京全式金公司市售产品。
PCR模板DNA：土壤宏基因组DNA
二、实验方法
1.根据测序获得的宏基因组基因序列设计1对PCR特异性引物：
dicX4-F：5′ACCACTAGTATGCCTTTCGTTTACAATGC 3′
dicX4-R：5′ACCCATATGTTACCCTCTCAATCCGGTGC 3′
2.通过PCR方法从宏基因组DNA中扩增出目的基因序列。
反应条件：95℃ 10min，[95℃ 30sec，61℃ 30sec，72℃ 1.0min]35个循环，72℃ 10min。
3.PCR产物经胶回收后，克隆于载体pJET上，命名为pJET-dicX4，并测序验证；然后通过Spe I/Nde I双酶切获得含有粘性末端的dicX4基因及含有groEL启动子的pRAD1载体，构建大肠杆菌表达载体pRAD-dicX4，将该表达载体转化大肠杆菌JM109，经PCR、酶切，测序验证插入序列正确，将该菌株命名为JM-dicX4。含有pRAD1对照空质粒的E.coli JM109命名为JM-D1。
三、实验结果
利用PCR技术成功克隆了土壤宏基因组中dicX4基因，成功构建了表达dicX4基因的重组大肠杆菌工程菌株。经PCR、酶切、测序验证插入序列正确，将该菌株命名为JM-dicX4。含有pRAD1对照空质粒的E.coli JM109，命名为JM-D1。
四、实验结论
完成表达dicX4的重组大肠杆菌工程菌株的构建。
实施例2 表达dicX4的重组大肠杆菌工程菌株的麦草畏代谢能力分析
一、实验材料
重组工程菌株：实施例1得到的表达dicX4基因的JM-dicX4菌株
对照菌株：实施例1所述含空质粒的JM-D1菌株。
二、实验方法
1.将对照菌株和重组工程菌株在LB固体培养基平板上划线活化；
2.挑取单菌落接种于添加有相应抗生素的液体LB培养基中，37℃培养至指数生长中后期；
3. 4000rpm,4min,离心收集菌体，用MSM培养基重悬、洗涤菌体两遍；
4.将菌株重悬于含有500mg/L麦草畏的50mL MSM培养基中；
5.分别在0h、12h、24h、36h、48h和60h等不同时间点收菌，测定OD 600，并观察培养液颜色与浑浊度。
三、实验结果
如图1所示，重组工程菌JM-dicX4能够代谢麦草畏。图1中表达dicX4基因的重组工程菌JM-dicX4能够在以麦草畏为唯一碳源的培养基中生长，培养基变浑浊，48h孵育培养后，麦草畏被降解，培养基颜色逐渐变红。图1含有空质粒的对照菌株JM-D1菌株孵育48h后，培养基依然澄清，未见颜色发生变化。
四、实验结论
表达dicX4基因的重组工程菌JM-dicX4能够在以麦草畏为唯一碳源的培养基中生长，具有降解除草剂麦草畏的能力。
实施例3 表达dicX4的重组大肠杆菌工程菌株的催化活性实验
一、实验材料
重组工程菌株：实施例1得到的表达dicX4基因的JM-dicX4菌株
对照菌株：实施例1所述含空质粒的JM-D1菌株。
二、实验方法
1.将对照菌株和重组工程菌株在LB固体培养基平板上划线活化；
2.挑取单菌落接种于添加有相应抗生素的液体LB培养基中，37℃培养至指数生长中后期；
3. 4000rpm,4min,离心收集菌体，用MSM培养基重悬、洗涤菌体两遍；
4.将菌株重悬于含有500mg/L麦草畏的50mL MSM培养基中；
5.分别在0h、24h、48h和60h等不同时间点收菌，测定OD 600，并取样用于HPLC测定培养液中麦草畏的含量。
利用惠普1050系列HPLC系统进行高效液相色谱分析。色谱柱为C18柱。样品上样量为100μL，麦草畏及其降解产物利用移动相的梯度变化进行稀释分析。移动相溶液B变化梯度为30min中从30％-95％。(A：超纯水：乙腈∶甲醇：乙酸＝58.4:31.7:7.5:2.4；B：100％乙腈)，流速为0.8mL/min。检测波长：麦草畏λ＝275nm；DCSA λ＝319nm。
三、实验结果
如图2所示，A中除草剂麦草畏的紫外检测波长为λ＝275nm，出峰时间约6min处；B中其降解产物DCSA峰为紫外检测波长为λ＝319nm，出峰时间约5.1min处。
如图3所示，表达dicX4基因的重组工程菌JM-dicX4与含有空质粒的对照菌株JM-D1菌株产物峰型显著不同。图3A中，48h孵育培养后，对照菌株JM-D1(虚线)的培养基中的主要化合物峰仍为1号峰，保留时间在6min处，对照标准品结果该峰为麦草畏化合物。而图3A中，表达dicX4基因的重组工程菌JM-dicX4(实线)培养基中的化合物峰型发生了明显的变化，1号峰已经检测不到。图3B中显示，48h孵育培养后，重组工程菌JM-dicX4(实线)的主要产物为2号峰，保留时间在5.1min处，对照标准品结果该峰为DCSA化合物。而对照菌株JM-D1(虚线)未检测到2号峰物质产生。
四、实验结论
重组表达的DicX4蛋白质具有降解除草剂麦草畏的催化功能，降解产物主要为无除草剂活性的3,6-二氯水杨酸(DCSA)。该基因在抗除草剂转基因农作物的培育及污染土壤的生物修复方面具有应用潜力。

Claims (6)

  1. SEQ ID NO:1所示序列的基因。
  2. 权利要求1所述基因的应用，是作为除草剂降解基因在抗除草剂转基因作物培育和环境生物修复降解除草剂中的应用。
  3. 权利要求2所述的应用，所述降解除草剂是将有毒性的麦草畏降解为无毒性物质的反应。
  4. 含有SEQ ID NO:1所示序列基因的质粒。
  5. 权利要求4所述的质粒在抗除草剂转基因作物培育及除草剂降解中的应用。
  6. 权利要求1所述DNA序列编码的氨基酸序列，如SEQ ID NO:2所示。
PCT/CN2020/108309 2019-10-14 2020-08-11 具有除草剂麦草畏降解功能的基因dicX4及其应用 WO2021073220A1 (zh)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
CN201910977573.2 2019-10-14
CN201910977573.2A CN111118037B (zh) 2019-10-14 2019-10-14 具有除草剂麦草畏降解功能的基因dicx4及其应用

Publications (1)

Publication Number Publication Date
WO2021073220A1 true WO2021073220A1 (zh) 2021-04-22

Family

ID=70495336

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/CN2020/108309 WO2021073220A1 (zh) 2019-10-14 2020-08-11 具有除草剂麦草畏降解功能的基因dicX4及其应用

Country Status (2)

Country Link
CN (1) CN111118037B (zh)
WO (1) WO2021073220A1 (zh)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117286161A (zh) * 2023-11-27 2023-12-26 南京农业大学三亚研究院 麦草畏厌氧降解中间产物3，6-二氯水杨酸脱羧酶CsaDC及其应用

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111118037B (zh) * 2019-10-14 2021-07-13 中国农业科学院生物技术研究所 具有除草剂麦草畏降解功能的基因dicx4及其应用

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002068607A2 (en) * 1997-04-04 2002-09-06 Board Of Regents Of The University Of Nebraska Methods and materials for making and using transgenic dicamba-degrading organisms
CN109439674A (zh) * 2018-12-07 2019-03-08 南京农业大学 一种通过转dmo基因提高大豆抗除草剂麦草畏和植物感光性的方法
CN111118037A (zh) * 2019-10-14 2020-05-08 中国农业科学院生物技术研究所 具有除草剂麦草畏降解功能的基因dicx4及其应用

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998045424A1 (en) * 1997-04-04 1998-10-15 Weeks Donald P Methods and materials for making and using transgenic dicamba-degrading organisms
US7838729B2 (en) * 2007-02-26 2010-11-23 Monsanto Technology Llc Chloroplast transit peptides for efficient targeting of DMO and uses thereof
CN105802933B (zh) * 2016-03-22 2020-05-05 北京大北农科技集团股份有限公司 除草剂耐受性蛋白质、其编码基因及用途
CN107794271B (zh) * 2017-11-03 2020-06-23 南京农业大学 一个龙胆酸双加氧酶及其编码基因和应用
CN109971773B (zh) * 2019-03-29 2022-02-22 南阳师范学院 一种编码可降解3-氯龙胆酸的龙胆酸双加氧酶DsmI的基因dsmI及其应用

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2002068607A2 (en) * 1997-04-04 2002-09-06 Board Of Regents Of The University Of Nebraska Methods and materials for making and using transgenic dicamba-degrading organisms
CN109439674A (zh) * 2018-12-07 2019-03-08 南京农业大学 一种通过转dmo基因提高大豆抗除草剂麦草畏和植物感光性的方法
CN111118037A (zh) * 2019-10-14 2020-05-08 中国农业科学院生物技术研究所 具有除草剂麦草畏降解功能的基因dicx4及其应用

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN117286161A (zh) * 2023-11-27 2023-12-26 南京农业大学三亚研究院 麦草畏厌氧降解中间产物3，6-二氯水杨酸脱羧酶CsaDC及其应用
CN117286161B (zh) * 2023-11-27 2024-03-19 南京农业大学三亚研究院 麦草畏厌氧降解中间产物3，6-二氯水杨酸脱羧酶CsaDC及其应用

Also Published As

Publication number Publication date
CN111118037A (zh) 2020-05-08
CN111118037B (zh) 2021-07-13

Similar Documents

Publication Publication Date Title
CN109666658B (zh) 用于制备nmn的烟酰胺磷酸核糖转移酶、编码基因、重组载体及应用
Liu et al. Iterative genome editing of Escherichia coli for 3-hydroxypropionic acid production
WO2021184557A1 (zh) 一种生物多酶偶联法氧化还原不对称制备l-草铵膦的方法
WO2021073220A1 (zh) 具有除草剂麦草畏降解功能的基因dicX4及其应用
CN107794271B (zh) 一个龙胆酸双加氧酶及其编码基因和应用
CN109971773B (zh) 一种编码可降解3-氯龙胆酸的龙胆酸双加氧酶DsmI的基因dsmI及其应用
CN110373370B (zh) 一种耦合atp再生系统的催化体系及其在生产谷胱甘肽过程中的应用
CN106701698A (zh) 羰基还原酶、突变体及其在制备抗真菌类药物中间体中的应用
CN114450403A (zh) 用于在枫糖尿症(msud)的治疗中使用的酶的生物合成
US10787489B2 (en) Biocatalyst comprising photoautotrophic organisms producing recombinant enzyme for degradation of harmful algal bloom toxins
CN106995794B (zh) 一种提高丁二酸产量的产琥珀酸放线杆菌工程菌株及其构建方法与用途
CN113430181B (zh) 一种来源亚洲象肠道宏基因组的细菌漆酶及其基因
US20110207190A1 (en) Methods of xylitol preparation
CN111321092B (zh) (r,s)-2,4-滴丙酸降解菌株及特异性降解其(s)-型异构体的双加氧酶基因
US10934521B2 (en) Genetically modified microorganisms
CN108441503B (zh) 麦草畏中间产物3,6-二氯龙胆酸脱氯酶DsmH2及其编码基因的应用
CN110872591B (zh) 除草剂麦草畏降解基因dicX1及其应用
Xiong et al. High efficiency and throughput system in directed evolution in vitro of reporter gene
CN111139248B (zh) dicX5基因及其对除草剂的降解功能
CN111139249B (zh) 运动金球菌麦草畏降解基因dicM的用途
CN101177686B (zh) 一种丙酮酸氧化酶基因、其重组表达质粒及转化的菌株
CN111139238B (zh) 除草剂麦草畏的降解基因dicX3及其应用
CN107619832B (zh) 一种氯代硝基苯酚类化合物氧化还原酶基因簇cnpAB及其应用
CN107523580B (zh) 一种卤代对羟基苯甲酸氧化脱羧酶基因odcA及其应用
CN109112117B (zh) 一种分离的二化螟cyp15c1基因及其编码蛋白

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: 20877630

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 20877630

Country of ref document: EP

Kind code of ref document: A1