PLANTA
·
2017
·Vol. 246
·期 2
·277-280
·
IF 3.361
Bashandy Hany,Teeri Teemu H.
Main conclusion
Unauthorized genetically engineered orange petunias were found on the market.
Genetic engineering of petunia was shown to lead to novel flower color some 20 years ago. Here we show that petunia lines with orange flowers, generated for scientific purposes, apparently found their way to petunia breeding programmes, intentionally or unintentionally. Today they are widely available, but have not been registered for commerce.
研究领域:
植物学
·
遗传学
✓ 已验证
DOI
PubMed
高影响因子
TALANTA
·
2014
·
IF 3.511
Xiaohua Jiang,Huimin Zhang,Jun Wu,Xiang Yang,Jingwei Shao,Yujing Lu,Bin Qiu,Zhenyu Lin,Guonan Chen
In this paper, a novel label-free G-quadruplex DNAzyme sensor has been proposed for colorimetric identification of GMO using CaMV 35S promoter sequence as the target. The binary probes can fold into G-quadruplex structure in the presence of DNA-T (Target DNA) and then combine with hemin to form a DNAzyme resembling horseradish peroxidase . The detection system consists of two G-rich probes with 2:2 split mode by using the absorbance and color of ABTS 2− as signal reporter. Upon the addition of a target sequence, two probes both hybridize with target and then their G-rich sequences combine to form a G-quadruplex DNAzyme, and the DNAzyme can catalyze the reaction of ABTS 2− with H 2 O 2 . Then the linear range is from 0.05 to 0.5 μM while detection limit is 5 nM. These results demonstrate that the proposed G-quadruplex DNAzyme method could be used as a simple, sensitive and cost-effective approach for assays of GMO.
研究领域:
生物化学
·
分子生物学
·
食品科学
✓ 已验证
DOI
高影响因子
Scientific Reports
·
2014
·Vol. 4
·期 1
·1-8
·
IF 5.078
Zhou Dinggang,Guo Jinlong,Xu Liping,Gao Shiwu,Lin Qingliang,Wu Qibin,Wu Luguang,Que Youxiong
To meet the demand for detection of foreign genes in genetically modified (GM) sugarcane necessary for regulation of gene technology, an efficient method with high specificity and rapidity was developed for the cry1Ac gene, based on loop-mediated isothermal amplification (LAMP). A set of four primers was designed using the sequence of cry1Ac along with optimized reaction conditions: 5.25 mM of Mg 2+ , 4:1 ratio of inner primer to outer primer, 2.0 U of Bst DNA polymerase in a reaction volume of 25.0 μL. Three post-LAMP detection methods (precipitation, calcein (0.60 mM) with Mn 2+ (0.05 mM) complex and SYBR Green I visualization), were shown to be effective. The sensitivity of the LAMP method was tenfold higher than that of conventional PCR when using templates of the recombinant cry1Ac plasmid or genomic DNA from cry1Ac transgenic sugarcane plants. More importantly, this system allowed detection of the foreign gene on-site when screening GM sugarcane without complex and expensive instruments, using the naked eye. This method can not only provide technological support for detection of cry1Ac , but can also further facilitate the use of this detection technique for other transgenes in GM sugarcane.
研究领域:
分子生物学
·
植物生物技术
·
基因工程
✓ 已验证
产品关联
DOI
PubMed
高影响因子
MICROCHEMICAL JOURNAL
·
2021
·
IF 4.821
Liyuan Zhang,Runzhong Yu,Yingbo Yu
The gas chromatography-mass spectrometry was used to isolate and identify metabolites of non-transgenic and Bacillus thuringiensis transgenic maize. The non-targeted metabolomics technique was used to analysis the metabolic pathway and mechanism of two kinds of maize. The methanol was used as extractant and the N,O-bis(trimethylsilyl) trifluoroacetamide was used as derivatization reagent. 38 kinds of metabolites were isolated and identified from non-transgenic maize, and 61 kinds of metabolites were isolated and identified in Bacillus thuringiensis transgenic maize. The specific metabolites between non-transgenic and Bacillus thuringiensis transgenic maize were analyzed. The metabolic pathway of specific metabolites was analyzed by KEGG annotation. The metabolic mechanism of non-transgenic maize and Bacillus thuringiensis transgenic maize was explored. The result indicated there were more metabolites involved in metabolic pathways in Bacillus thuringiensis transgenic maize than in non-transgenic maize, and tricarboxylic acid cycle and energy metabolism pathways of Bacillus thuringiensis transgenic maize are found to be higher than that of non-transgenic maize. The metabolic pathway of Bacillus thuringiensis transgenic maize conforms to the biological activity law.
研究领域:
植物科学
·
分子生物学
·
代谢组学
✓ 已验证
产品关联
DOI
高影响因子
NUCLEIC ACIDS RESEARCH
·
2025
·Vol. 53
·期 2
·
IF 13.1
Zhang Lihang,Qiu Xinyuan,Zhou Yuting,Luo Zhengyang,Zhu Lingyun,Shao Jiawei,Xie Mingqi,Wang Hui
The CRISPR-derived endoribonuclease Csy4 is a popular tool for controlling transgene expression in various therapeutically relevant settings, but adverse effects potentially arising from non-specific RNA cleavage remains largely unexplored. Here, we report a split-Csy4 architecture that was carefully optimized for in vivo usage. First, we separated Csy4 into two independent protein moieties whose full catalytic activity can be restored via various constitutive or conditional protein dimerization systems. Next, we show that introduction of split-Csy4 into human cells caused a substantially reduced extent in perturbation of the endogenous transcriptome when directly compared to full-length Csy4. Inspired by these results, we went on to use such split-Csy4 module to engineer inducible CRISPR- and translation-level gene switches regulated by the FDA-approved drug grazoprevir. This work provides valuable resource for Csy4-related biomedical research and discusses important issues for the development of clinically eligible regulation tools.
研究领域:
分子生物学
·
基因工程
·
治疗学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
PLANT CELL AND ENVIRONMENT
·
2021
·Vol. 44
·期 9
·3173-3183
·
IF 7.228
Kunlong Su,Zhenying Wu,Yuchen Liu,Shanshan Jiang,Dongmei Ma,Yan Wang,Chunxiang Fu
Dinitrotoluene (DNT) has been extensively used in manufacturing munitions, polyurethane foams and other important chemical products. However, it is highly toxic and mutagenic to most organisms. Here, we synthesized a codon-optimized bacterial nitroreductase gene, NfsI, for plant expression. The kinetic analysis indicates that the recombinant NfsI can detoxify both 2,4-DNT and its sulfonate (DNTS), while it has a 97.6-fold higher catalytic efficiency for 2,4-DNT than DNTS. Furthermore, we overexpressed NfsI in switchgrass ( Panicum virgatum L.), which is a multiple-purpose crop used for fodder and biofuel production as well as phytoremediation. The 2,4-DNT treatment inhibited root elongation of wild-type switchgrass plants and promoted reactive oxygen species (ROS) accumulation in roots. In contrast, overexpression of NfsI in switchgrass significantly alleviated 2,4-DNT-induced root growth inhibition and ROS overproduction. Thus, the NfsI overexpressing transgenic switchgrass plant removed 94.1% 2,4-DNT after 6 days, whose efficiency was 1.7-fold higher than control plants. Moreover, the comparative transcriptome analysis suggests that 22.9% of differentially expressed genes induced by 2,4-DNT may participate in NfsI-mediated 2,4-DNT detoxification in switchgrass. Our work sheds light on the function of NfsI during DNT phytoremediation for the first time, revealing the application potential of switchgrass plants engineered with NfsI .
研究领域:
植物学
·
环境科学
·
遗传工程
✓ 已验证
DOI
高影响因子
Biomaterials Science
·
2017
·Vol. 5
·期 7
·1279-1292
·
IF 4.21
Yawen Liu,Zhaozhu Zheng,He Gong,Meng Liu,Shaozhe Guo,Gang Li,Xiaoqin Wang,David L. Kaplan
The structure of DNA is susceptible to alterations at high temperature and on changing pH, irradiation and exposure to DNase. Options to protect and preserve DNA during storage are important for applications in genetic diagnosis, identity authentication, drug development and bioresearch. In the present study, the stability of total DNA purified from human dermal fibroblast cells, as well as that of plasmid DNA, was studied in silk protein materials. The DNA/silk mixtures were stabilized on filter paper (silk/DNA + filter) or filter paper pre-coated with silk and treated with methanol (silk/DNA + PT-filter) as a route to practical utility. After air-drying and water extraction, 50–70% of the DNA and silk could be retrieved and showed a single band on electrophoretic gels. 6% silk/DNA + PT-filter samples provided improved stability in comparison with 3% silk/DNA + filter samples and DNA + filter samples for DNA preservation, with ∼40% of the band intensity remaining at 37 °C after 40 days and ∼10% after exposure to UV light for 10 hours. Quantitative analysis using the PicoGreen assay confirmed the results. The use of Tris/borate/EDTA (TBE) buffer enhanced the preservation and/or extraction of the DNA. The DNA extracted after storage maintained integrity and function based on serving as a functional template for PCR amplification of the gene for zinc finger protein 750 (ZNF750) and for transgene expression of red fluorescence protein (dsRed) in HEK293 cells. The high molecular weight and high content of a crystalline beta-sheet structure formed on the coated surfaces likely accounted for the preservation effects observed for the silk/DNA + PT-filter samples. Although similar preservation effects were also obtained for lyophilized silk/DNA samples, the rapid and simple processing available with the silk–DNA–filter membrane system makes it appealing for future applications.
研究领域:
遗传学
·
生物技术
·
材料科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
BIOSENSORS & BIOELECTRONICS
·
2017
·
IF 7.78
Hongfei Gao,Luke Wen,Yuhua Wu,Zhifeng Fu,Gang Wu
The development of genetically modified (GM) insect-resistant crops has aroused great public concern about the risks on the eco-environment resulting from a release of toxic Cry proteins (such as Cry1Ab) to the soil. Therefore, it is of crucial importance to measure the Cry proteins level and the GM crops content. Here, we have tested for the first time a method that uses novel carbon nanospheres (CNPs) label-free electrochemiluminescent (ECL) immunosensor for the ultrasensitive quantification of Cry1Ab and GM crops. In this work, novel CNPs were prepared from printer toner with a very facile approach, and linked with anti-Cry1Ab antibodies to modify a golden working electrode . The immunoreaction between Cry1Ab and its antibody formed an immunocomplex on the bioreceptor region of the sensor, which inhibited electron transfer between the electrode surface and the ECL substance, leading to a decrease of ECL response. Under the optimal conditions, the fabricated label-free ECL immunosensor determined Cry1Ab down to 3.0 pg mL −1 within a linear range of 0.010−1.0 ng mL −1 , showing significant improvement of sensitivity than that of most previous reports. Meanwhile, the proposed method was successfully applied for GM rice BT63 and GM maize MON810 detections down to 0.010% and 0.020%, respectively. Due to its outstanding advantages such as high sensitivity, ideal selectivity, simple fabrication, rapid detection, and low cost, the developed method can be considered as a powerful and pioneering tool for GM crops detection. Its use can also be extended to other toxin protein sensing in foods.
研究领域:
生物技术
·
环境科学
·
农业科学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
JOURNAL OF FOOD COMPOSITION AND ANALYSIS
·
2021
·
IF 4.556
Yue-Hong Pang,Yi-Ying Wang,Meng-Meng Sun,Xiao-Fang Shen
The cauliflower mosaicvirus 35S (CaMV35S), as the most common promoter, has a usage rate of 70 % in transgenic crops, especially in dicots. Herein, a visual detection method of rolling circle amplification (RCA) combined with DNAzyme technology was developed for the determination of CaMV35S gene. A template sequence of the G-quadruple gene was designed for combining with the target gene CaMV35S. When the target gene existed, the RCA reaction occurred through the action of the enzyme, and the product combined with hemin to generate DNAzyme, which catalyzed the color reaction of 2′-hydrazine-bis-3-ethylbenzothiazolin-6-sulfonic acid (ABTS). Under the optimal experimental conditions, the linear range is 1 × 10 −13 ∼1 × 10 -8 mol/L, and the detection limit is 3 × 10 -14 mol/L (S/N = 3). The method has been successfully applied to standard samples and practical samples (soybean and tofu). For verifying the feasibility and reliability, real-time fluorescent quantitative PCR method was used to detect the target gene CaMV35S, and the results were in good agreement with those of our method, indicating that our developed method can be used for the determination of actual samples.
研究领域:
分子生物学
·
基因工程
·
农业科学
✓ 已验证
产品关联
DOI
高影响因子
Analytical Methods
·
2014
·Vol. 6
·期 23
·9198-9201
·
IF 1.938
Rui Wang,Jian Wu,Fang Zhang,Liu Wang,Feng Ji
1% (w/v) concentration of pullulan as a cross-priming isothermal amplification (CPA) reaction additive has a great acceleration effect on nucleic acid amplification. Using this property of pullulan, on-point specific detection of nucleic acid by lateral-flow dipstick (LFD) within 20 minutes was achieved with initial DNA template of 750 copies.
研究领域:
分子生物学
·
食品科学
·
植物遗传学
✓ 已验证
产品关联
DOI
高影响因子
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
·
2024
·
IF 5.7
Xiao Han,Minghui Lu,Yaru Zhang,Xinru Liu,Qiang Zhang,Xue Bai,Shuli Man,Liangjuan Zhao,Long Ma
Genetically modified crops (GMCs) have been discussed due to unknown safety, and thus, it is imperative to develop an effective detection technology. CRISPR/Cas is deemed a burgeoning technology for nucleic acid detection. Herein, we developed a novel detection method for the first time, which combined thermostable Cas12b with loop-mediated isothermal amplification (LAMP), to detect genetically modified (GM) soybeans in a customized one-pot vessel. In our method, LAMP-specific primers were used to amplify the cauliflower mosaic virus 35S promoter (CaMV35S) of the GM soybean samples. The corresponding amplicons activated the trans-cleavage activity of Cas12b, which resulted in the change of fluorescence intensity. The proposed bioassay was capable of detecting synthetic plasmid DNA samples down to 10 copies/μL, and as few as 0.05% transgenic contents could be detected in less than 40 min. This work presented an original detection method for GMCs, which performed rapid, on-site, and deployable detection.
研究领域:
植物学
·
分子生物学
·
生物技术
✓ 已验证
DOI
高影响因子
Journal of Orthopaedic Translation
·
2024
·
IF 5.9
Yuejun Lin,Zhaowei Jiang,Jiaming Yang,Ming Wang,Haixing Wang,Xiaoting Zhang,Xuan Lu,Shanshan Bai,Tongzhou Liang,Botai Li,Jie Shao,Lu Zhang,Dashuang Gao,Jiajun Chen,Sien Lin,Fan Yang,Gang Li
Objective Distraction osteogenesis (DO) has been widely used to treat bone defects as its effectiveness in bone regeneration. Currently, distraction devices for establishing DO models are mainly developed for rats or large animals. However, a mouse DO model is in great need for in-depth mechanistic investigations using various transgenic mice. The current study reports the development of a reproducible murine DO model. Methods A mini-titanium lengthener was designed and fabricated. The mini-lengthener was applied on the murine femur with four threaded pins using a designed clamp as the drilling and insertion guide. After transverse osteotomy using a Gigli saw, and after 5 days of latency, DO procedures started at 0.3 mm/day for 10 days, and the consolidation period was left for 28 days. The bone formation was monitored by radiography and histology. Potential effects on animal locomotion during DO were also measured by behavior tests. Results Separated bone segments maintained good alignment during the entire DO phases. New bone formation was found as early as the end of the distraction phase. Active bone remodeling was found between the separated bone segments at late distraction and early consolidation phases. At the mature consolidation phase, bone remodeling was mainly observed in the contact cortical bone. Mice underwent DO procedure did not have significant impairment in their locomotion. Conclusion We have successfully developed a murine femoral DO model, which may be used to study the biological processes of DO. We also developed the mini-lengthener and the guide clamp to ensure the standardization and reproducibility of the mouse DO model. The translational potential of this article: Current study reports the development of a murine femoral DO model. A well-established murine DO model will facilitate further investigations of the biological mechanisms of DO in various transgenic and normal mice.
研究领域:
骨科
·
再生医学
·
生物力学
✓ 已验证
产品关联
DOI
PubMed
高影响因子
危险品化学品经营许可证（带存储）