JOURNAL OF FLUORESCENCE
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2015
·Vol. 26
·期 2
·451-457
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IF 1.927
Liu Cui,Zhou Zhen,Zou Linling,Cao Yuan-Cheng,Liu Jun’An,Lin Yongjun
A highly sensitive transgenic protein analysis method was proposed here based on fluorescein diacetate (FDA). First, FDA was prepared by the ball mill to harvest the nano-sized organic particles. Further examines showed that the FDA size can be controlled by the speed of centrifugation which can obtain FDA in well-distributed size. Cy3 antibody immobilization tests showed that the proteins can attach onto the FDA particles while keep bioactivities. FDA and Cry1Ac antibody immunoassay tests showed that when the FDA particle was in 150 nm, the linear range was 0.01 ng/L-30 μg/mL. And it has the lower detection limitation of 0.01 ng/L, which is 100 times more sensitive than the ELISA methods. These results indicate that the FDA related immunoassays are the promising approach in the transgenic analysis.
研究领域:
生物化学
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分子生物学
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食品科学
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PubMed
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
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植物生物技术
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基因工程
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PubMed
高影响因子
Insect Science
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2020
·Vol. 28
·期 6
·1756-1765
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IF 2.791
Shan Wang,Min Zhang,Jinyong Huang,Leyao Li,Kaiyuan Huang,Yuting Zhang,Yalu Li,Zhongyuan Deng,Xinzhi Ni,Xianchun Li
Genetically engineered crops simultaneously produce defensive allelochemicals and Bacillus thuringiensis (Bt) toxin proteins to kill some of the world's most devastating insect pests. How the two types of toxins, when ingested sequentially or simultaneously, interact at both lethal and sublethal doses in these pests remains underexplored. Here, we examined the toxicological interactions between the Bt toxin Cry1Ac and the flavonoid allelochemical flavone in Helicoverpa armigera . Simultaneous exposure of H. armigera neonates to lethal doses (LC 25 ) of Cry1Ac and flavone caused a mortality significantly higher than that of either toxin alone and their expected additive mortality. Preexposure for 24 h to a sublethal dose (LC 10 ) of Cry1Ac followed by 6-d simultaneous exposure to the same dose of Cry1Ac plus a lethal dose (1.6 mg/g diets, LC 50 ) of flavone resulted in a mortality significantly higher than that of the LC 50 dose of flavone alone and the expected additive mortality of the LC 50 dose of flavone plus the LC 10 dose of Cry1Ac. One-day preexposure to the sublethal dose (LC 10 ) of flavone followed by 6-d simultaneous exposure to the LC 50 dose (6 ng/cm 2 ) of Cry1Ac plus the LC 10 dose of flavone yielded a mortality significantly higher than that of the LC 50 dose of Cry1Ac but similar to the expected additive mortality of the LC 50 dose of Cry1Ac plus the LC 10 dose of flavone. The results suggest that Cry1Ac induces and synergizes the toxicity of flavone against H. armigera larvae.
研究领域:
植物学
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昆虫学
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遗传工程
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高影响因子
SENSORS AND ACTUATORS B-CHEMICAL
·
2016
·
IF 4.758
Jiyuan Liang,Yuxiang Wu,Cui Liu,Yuan-Cheng Cao,Jun’An Liu,Yongjun Lin
The silicon coated Magnetic nanoparticles (MNps) were prepared and applied to the genetically modified detection for Cry1Ac protein. TEM observations showed the bare MNps were in the size of 3–5 nm and the silicon coated core/shell MNps were in the range of 30–60 nm 1 M HCl acidic solution was used to test the chemical stability of the resultant MNps and results showed the coated MNps were very stable in the acid environment even after 12 h. Cy3 labelled IgG tests showed that the probe on the MNps was kept the bioactivity in the immunoassays. MNps based detection results showed that this analytical strategy had similar linear relation which R = 0.995 with the commercial ELISA kit. The MNps based detections were reliable for the Cry1 Ac protein in the linear range from 0.1 pg/L to 1.0 mg/L, and showed better performance in the low concentration of analyte. This study provides a promising technique to detect the Cry1Ac protein with high sensitivity in the examination of the GM products which is of importance in food safety.
研究领域:
生物技术
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食品安全
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纳米科技
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DOI
高影响因子
PEST MANAGEMENT SCIENCE
·
2025
·
IF 3.8
Cheng Shen,Guochen Li,Tianyi Han,Naila Ilyas,Meng He,Ziyan Huang,Xiao Zhang,Chongxin Xu,Chengyu Chen,Wei Chen,Xianjin Liu,Qing Zhu,Yuan Liu
BACKGROUND Bacillus thuringiensis (Bt) toxins are widely used in pest control owing to their effectiveness as bioinsecticides. However, the extensive use of these toxins has resulted in the development of resistance in various pest populations, including Plutella xylostella , which is a significant pest of cruciferous crops. This increasing resistance highlights the need for alternative insecticidal strategies that can complement or replace traditional methods. RESULTS A novel approach involves using insect cadherin receptors to screen for nanobodies with potential insecticidal activity that selectively target the toxin binding region of the Plutella xylostella cadherin toxin via phage display technology. Through three rounds of panning with a natural alpaca nanobody phage display library, we identified nanobody B1, which specifically binds to the CR9-CR11 region of Plutella xylostella cadherin. This nanobody significantly inhibited the binding of Cry1Ac toxin to cadherin, a key factor in insect susceptibility to the toxin. In bioassays, the application of soluble B1 protein (80 μg/cm 2 ) resulted in 25 ± 4.5% mortality in Plutella xylostella larvae. Histological examination of the midgut tissue revealed substantial damage to the epithelial layer, supporting the bioassay findings. Enzymatic hydrolysis assays demonstrated that, while B1 exhibited some stability, it showed limited resistance to enzymatic degradation in vitro . This could be a significant contributing factor to the low insecticidal activity of B1. CONCLUSION This study proposes a novel strategy for insecticide development based on the application of nanobodies to target cadherins within the midgut of Plutella xylostella . The identification of nanobody B1 presents a novel approach to circumvent resistance mechanisms in pest populations, overcoming resistance mechanisms in pest populations, and thus advancing sustainable pest management practices. Furthermore, these findings establish a foundation for designing alternative insecticidal proteins that could diminish the reliance on traditional formulations. © 2025 Society of Chemical Industry.
研究领域:
昆虫学
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生物技术
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害虫防治
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分子生物学
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生物化学
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DOI
PubMed
高影响因子
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