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https://www.readbyqxmd.com/read/28429650/development-of-real-time-pcr-tests-for-the-detection-of-tenebrio-molitor-in-food-and-feed
#1
Frédéric Debode, Aline Marien, Amaury Gerard, Frédéric Francis, Olivier Fumiere, Gilbert Berben
Insects are rich in proteins and could be an alternative source of proteins to feed animals and humans. Numerous companies have started the production of insects for feed purposes. In Europe, these processed animal proteins are not yet authorized by legislation, as many questions still need to be answered concerning this "novel food". Authorizations will be possible when methods of authentication of the products are available. In this study we propose real-time PCR methods for the specific detection of the mealworm (Tenebrio molitor), one of the most widely used insects for food and feed production...
April 21, 2017: Food Additives & Contaminants. Part A, Chemistry, Analysis, Control, Exposure & Risk Assessment
https://www.readbyqxmd.com/read/28389549/identification-of-holotrichia-oblita-midgut-proteins-that-bind-to-cry8-like-toxin-from-bacillus-thuringiensis-and-assembling-of-h-oblita-midgut-tissue-transcriptome
#2
Jian Jiang, Ying Huang, Changlong Shu, Mario Soberón, Alejandra Bravo, Chunqing Liu, Fuping Song, Jinsheng Lai, Jie Zhang
The Bacillus thuringiensis strain HBF-18 (CGMCC2070), containing two cry genes (cry8-like and cry8Ga), is toxic to Holotrichia oblita larvae. Both Cry8-like and Cry8Ga proteins are active against this insect pest, while Cry8-like is the more toxic protein. To analyze the binding characteristics of Cry8-like and Cry8Ga proteins to brush border membrane vesicles (BBMV) in H. oblita larvae, binding assays were conducted with a fluorescent DyLight488-labeled Cry8-like toxin. Results of binding saturation assays demonstrated that Cry8-like binds specifically to binding sites on BBMV from H...
April 7, 2017: Applied and Environmental Microbiology
https://www.readbyqxmd.com/read/28247982/cytotoxicity-and-binding-profiles-of-activated-cry1ac-and-cry2ab-to-three-insect-cell-lines
#3
Jizhen Wei, Gemei Liang, Kongming Wu, Shaohua Gu, Yuyuan Guo, Xinzhi Ni, Xianchun Li
While Cry1Ac has been known to bind with larval midgut proteins cadherin, APN (amino peptidase N), ALP (alkaline phosphatase) and ABCC2 (ATP-binding cassette transporter subfamily C2), little is known about the receptors of Cry2Ab. To provide a clue to the receptors of Cry2Ab, we tested the baseline cytotoxicity of activated Cry1Ac and Cry2Ab against the midgut and fat body cell lines of Helicoverpa zea and the ovary cell line of Spodoptera frugiperda (SF9). As expected, the descending order of cytotoxicity of Cry1Ac against the three cell lines in terms of LC50 was midgut (31...
March 1, 2017: Insect Science
https://www.readbyqxmd.com/read/28220242/expression-of-clonorchis-sinensis-giiispla2-protein-in-baculovirus-infected-insect-cells-and-its-overexpression-facilitating-epithelial-mesenchymal-transition-in-huh7-cells-via-akt-pathway
#4
Mei Shang, Zhizhi Xie, Zeli Tang, Lei He, Xiaoyun Wang, Caiqin Wang, Yinjuan Wu, Ye Li, Lu Zhao, Zhiyue Lv, Zhongdao Wu, Yan Huang, Xinbing Yu, Xuerong Li
Although prior studies confirmed that group III secretory phospholipase A2 of Clonorchis sinensis (CsGIIIsPLA2) had stimulating effect on liver fibrosis by binding to LX-2 cells, large-scale expression of recombinant protein and its function in the progression of hepatoma are worth exploring. Because of high productivity and low lipopolysaccharides (LPS) in the Sf9-baculovirus expression system, we firstly used this system to express the coding region of CsGIIIsPLA2. The molecular weight of recombinant CsGIIIsPLA2 protein was about 34 kDa...
February 20, 2017: Parasitology Research
https://www.readbyqxmd.com/read/28108197/identification-of-aminopeptidase-n2-as-a-cry2ab-binding-protein-in-manduca-sexta
#5
Janette Onofre, Meztlli O Gaytán, Arlen Peña-Cardeña, Blanca I García-Gomez, Sabino Pacheco, Isabel Gómez, Alejandra Bravo, Mario Soberón
Bacillus thuringiensis Cry2Ab toxin has been used in combination with Cry1Ac for resistance management on the Bt-cotton that is widely planted worldwide. However, little is known regarding Cry2Ab mode of action. Particularly, there is a gap of knowledge on the identification of insect midgut proteins that bind Cry2Ab and mediate toxicity. In the case of Cry1Ab toxin, a transmembrane cadherin protein and glycosyl-phosphatidylinositol (GPI) anchored proteins like aminopeptidase-N1 (APN1) or alkaline-phosphatase (ALP) from Manduca sexta, have been shown to be important for oligomer formation and insertion into the membrane...
January 17, 2017: Peptides
https://www.readbyqxmd.com/read/27986933/the-spodoptera-exigua-lepidoptera-noctuidae-abcc2-mediates-cry1ac-cytotoxicity-and-in-conjunction-with-cadherin-contributes-to-enhance-cry1ca-toxicity-in-sf9-cells
#6
Xiang-Liang Ren, Wei-Li Jiang, Ya-Jie Ma, Hong-Yan Hu, Xiao-Yan Ma, Yan Ma, Guo-Qing Li
In insects, the mode of Cry1A toxins action has been studied in detail and many receptors that participate in the process are known. Recent evidence has revealed that an ABC transporter (ABCC2) is involved in conferring resistance to Cry1A toxins and that ABCC2 could be a receptor of Cry1A. However, it is not known whether Cry1Ca interacts with the same receptor proteins as Cry1A. In this study, we report the cloning of an ABC transporter gene, SeABCC2b, from the midgut of Spodoptera exigua (Hübner) larvae, and its expression in Sf9 cells for a functional analysis...
December 2016: Journal of Economic Entomology
https://www.readbyqxmd.com/read/27914919/expressing-a-moth-abcc2-gene-in-transgenic-drosophila-causes-susceptibility-to-bt-cry1ac-without-requiring-a-cadherin-like-protein-receptor
#7
Tristan Stevens, Sisi Song, John B Bruning, Amanda Choo, Simon W Baxter
Bt toxins ingested by insect pests can bind to midgut receptors and cause death, although several steps in this process remain unclear. Multiple Bt toxin receptors have been identified in Lepidoptera, including a cadherin-like protein (CaLP), which is central to several models explaining Bt toxins' mode of action. Mutations in the Plutella xylostella ATP-dependent binding cassette transporter C2 (Px-abcc2), rather than CaLP, are genetically linked with Bt Cry1Ac resistance. Here we expressed Px-abcc2 in Drosophila and performed larval bioassays to determine whether this protein acts as an effective Bt receptor...
January 2017: Insect Biochemistry and Molecular Biology
https://www.readbyqxmd.com/read/27867074/toxicity-of-cry1a-toxins-from-bacillus-thuringiensis-to-cf1-cells-does-not-involve-activation-of-adenylate-cyclase-pka-signaling-pathway
#8
Leivi Portugal, Carlos Muñóz-Garay, Diana L Martínez de Castro, Mario Soberón, Alejandra Bravo
Bacillus thuringiensis (Bt) bacteria produce Cry toxins that are able to kill insect pests. Different models explaining the mode of action of these toxins have been proposed. The pore formation model proposes that the toxin creates pores in the membrane of the larval midgut cells after interaction with different receptors such as cadherin, aminopeptidase N and alkaline phosphatase and that this pore formation activity is responsible for the toxicity of these proteins. The alternative model proposes that interaction with cadherin receptor triggers an intracellular cascade response involving protein G, adenylate cyclase (AC) and protein kinase A (PKA)...
January 2017: Insect Biochemistry and Molecular Biology
https://www.readbyqxmd.com/read/27813251/functional-characterization-of-bacillus-thuringiensis-cry-toxin-receptors-explains-resistance-in-insects
#9
Shiho Tanaka, Haruka Endo, Satomi Adegawa, Shingo Kikuta, Ryoichi Sato
Bacillus thuringiensis produces Cry toxins, which are used as insecticides in sprays and in transgenic crops. However, little is known about the function of Cry toxin receptors and the mechanisms that determine their binding specificity and activity. In this study, the cRNAs of Bombyx mori ABC transporter C2 (BmABCC2), the toxin-binding region of cadherin-like receptor (BtR175-TBR), or aminopeptidase N1 (BmAPN1) were injected into Xenopus oocytes, and the Cry1Aa-dependent cation-selective pore formation activities of these receptors were analyzed using a two-electrode voltage clamp...
December 2016: FEBS Journal
https://www.readbyqxmd.com/read/27792329/excess-imidacloprid-exposure-causes-the-heart-tube-malformation-of-chick-embryos
#10
Lin-Rui Gao, Shuai Li, Jing Zhang, Chang Liang, En-Ni Chen, Shi-Yao Zhang, Manli Chuai, Yong-Ping Bao, Guang Wang, Xuesong Yang
As a neonicotinoid pesticide, imidacloprid is widely used to control sucking insects on agricultural planting and fleas on domestic animals. However, the extent to which imidacloprid exposure has an influence on cardiogensis in early embryogenesis is still poorly understood. In vertebrates, the heart is the first organ to be formed. In this study, to address whether imidacloprid exposure affects early heart development, the early chick embryo has been used as an experimental model because of its accessibility at its early developmental stage...
November 30, 2016: Journal of Agricultural and Food Chemistry
https://www.readbyqxmd.com/read/27706708/characterization-of-a-novel-cry8ea3-binding-v-atpase-subunit-a-in-holotrichia-parallela
#11
W Wei, G Wei, Z Dan, Y Xiaoping, Z Yakun
Several receptor proteins of Cry toxin have been previously identified, including cadherin-like, aminopeptidase N, and alkaline phosphatase. In the present work, a novel binding protein, V-ATPase subunit A (HpVAA), was identified in Holotricia parallela larvae and characterized. We performed reverse transcription-polymerase chain reaction and rapid amplification of cDNA ends technology to obtain the cDNA of the full-length hpvaa. Sequencing analysis showed that the open reading frame of hpvaa (GenBank accession No...
September 2, 2016: Genetics and Molecular Research: GMR
https://www.readbyqxmd.com/read/27343383/functional-validation-of-cadherin-as-a-receptor-of-bt-toxin-cry1ac-in-helicoverpa-armigera-utilizing-the-crispr-cas9-system
#12
Jing Wang, Haonan Zhang, Huidong Wang, Shan Zhao, Yayun Zuo, Yihua Yang, Yidong Wu
Cadherins have been identified as receptors of Bacillus thuringiensis (Bt) Cry1A toxins in several lepidopteran insects including the cotton bollworm, Helicoverpa armigera. Disruption of the cadherin gene HaCad has been genetically linked to resistance to Bt toxin Cry1Ac in H. armigera. By using the CRISPR/Cas9 genome editing system (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated protein 9), HaCad from the Cry1Ac-susceptible SCD strain of H. armigera was successfully knocked out...
September 2016: Insect Biochemistry and Molecular Biology
https://www.readbyqxmd.com/read/27334721/rnai-induced-knockdown-of-a-cadherin-like-protein-ef531715-does-not-affect-toxicity-of-cry34-35ab1-or-cry3aa-to-diabrotica-virgifera-virgifera-larvae-coleoptera-chrysomelidae
#13
Sek Yee Tan, Murugesan Rangasamy, Haichuan Wang, Ana María Vélez, James Hasler, David McCaskill, Tao Xu, Hong Chen, Jessica Jurzenski, Matthew Kelker, Xiaoping Xu, Kenneth Narva, Blair D Siegfried
The western corn rootworm (WCR), Diabrotica virgifera virgifera LeConte, is an important maize pest throughout most of the U.S. Corn Belt. Bacillus thuringiensis (Bt) insecticidal proteins including modified Cry3Aa and Cry34/35Ab1 have been expressed in transgenic maize to protect against WCR feeding damage. To date, there is limited information regarding the WCR midgut target sites for these proteins. In this study, we examined whether a cadherin-like gene from Diabrotica virgifera virgifera (DvvCad; GenBank accession # EF531715) associated with WCR larval midgut tissue is necessary for Cry3Aa or Cry34/35Ab1 toxicity...
August 2016: Insect Biochemistry and Molecular Biology
https://www.readbyqxmd.com/read/27120167/continuous-evolution-of-bacillus-thuringiensis-toxins-overcomes-insect-resistance
#14
Ahmed H Badran, Victor M Guzov, Qing Huai, Melissa M Kemp, Prashanth Vishwanath, Wendy Kain, Autumn M Nance, Artem Evdokimov, Farhad Moshiri, Keith H Turner, Ping Wang, Thomas Malvar, David R Liu
The Bacillus thuringiensis δ-endotoxins (Bt toxins) are widely used insecticidal proteins in engineered crops that provide agricultural, economic, and environmental benefits. The development of insect resistance to Bt toxins endangers their long-term effectiveness. Here we have developed a phage-assisted continuous evolution selection that rapidly evolves high-affinity protein-protein interactions, and applied this system to evolve variants of the Bt toxin Cry1Ac that bind a cadherin-like receptor from the insect pest Trichoplusia ni (TnCAD) that is not natively bound by wild-type Cry1Ac...
May 5, 2016: Nature
https://www.readbyqxmd.com/read/26633693/binding-and-oligomerization-of-modified-and-native-bt-toxins-in-resistant-and-susceptible-pink-bollworm
#15
Josue Ocelotl, Jorge Sánchez, Raquel Arroyo, Blanca I García-Gómez, Isabel Gómez, Gopalan C Unnithan, Bruce E Tabashnik, Alejandra Bravo, Mario Soberón
Insecticidal proteins from Bacillus thuringiensis (Bt) are used extensively in sprays and transgenic crops for pest control, but their efficacy is reduced when pests evolve resistance. Better understanding of the mode of action of Bt toxins and the mechanisms of insect resistance is needed to enhance the durability of these important alternatives to conventional insecticides. Mode of action models agree that binding of Bt toxins to midgut proteins such as cadherin is essential for toxicity, but some details remain unresolved, such as the role of toxin oligomers...
2015: PloS One
https://www.readbyqxmd.com/read/26524384/a-high-throughput-in-vitro-assay-for-bacillus-thuringiensis-insecticidal-proteins
#16
Michi Izumi Willcoxon, Jaclyn R Dennis, Sabina I Lau, Weiping Xie, You You, Song Leng, Ryan C Fong, Takashi Yamamoto
A high-throughput, in-vitro assay for Bacillus thuringiensis (Bt) insecticidal proteins designated as Cry was developed and evaluated for screening a large number of Cry protein variants produced by DNA shuffling. This automation-amenable assay exploits an insect cell line expressing a single receptor of Bt Cry proteins. The Cry toxin used to develop this assay is a variant of the Cry1Ab protein called IP1-88, which was produced previously by DNA shuffling. Cell mortality caused by the activated Bt Cry toxin was determined by chemical cell viability assay in 96/384-well microtiter plates utilizing CellTiter 96(®) obtained from Promega...
January 10, 2016: Journal of Biotechnology
https://www.readbyqxmd.com/read/26240680/stem-cell-autotomy-and-niche-interaction-in-different-systems
#17
REVIEW
David C Dorn, August Dorn
The best known cases of cell autotomy are the formation of erythrocytes and thrombocytes (platelets) from progenitor cells that reside in special niches. Recently, autotomy of stem cells and its enigmatic interaction with the niche has been reported from male germline stem cells (GSCs) in several insect species. First described in lepidopterans, the silkmoth, followed by the gipsy moth and consecutively in hemipterans, foremost the milkweed bug. In both, moths and the milkweed bug, GSCs form finger-like projections toward the niche, the apical cells (homologs of the hub cells in Drosophila)...
July 26, 2015: World Journal of Stem Cells
https://www.readbyqxmd.com/read/26220297/transcriptional-analysis-of-susceptible-and-resistant-european-corn-borer-strains-and-their-response-to-cry1f-protoxin
#18
Neetha Nanoth Vellichirammal, Haichuan Wang, Seong-Il Eyun, Etsuko N Moriyama, Brad S Coates, Nicholas J Miller, Blair D Siegfried
BACKGROUND: Despite a number of recent reports of insect resistance to transgenic crops expressing insecticidal toxins from Bacillus thuringiensis (Bt), little is known about the mechanism of resistance to these toxins. The purpose of this study is to identify genes associated with the mechanism of Cry1F toxin resistance in European corn borer (Ostrinia nubilalis Hübner). For this, we compared the global transcriptomic response of laboratory selected resistant and susceptible O. nubilalis strain to Cry1F toxin...
2015: BMC Genomics
https://www.readbyqxmd.com/read/26160901/efficient-crispr-mediated-gene-targeting-and-transgene-replacement-in-the-beetle-tribolium-castaneum
#19
Anna F Gilles, Johannes B Schinko, Michalis Averof
Gene-editing techniques are revolutionizing the way we conduct genetics in many organisms. The CRISPR/Cas nuclease has emerged as a highly versatile, efficient and affordable tool for targeting chosen sites in the genome. Beyond its applications in established model organisms, CRISPR technology provides a platform for genetic intervention in a wide range of species, limited only by our ability to deliver it to cells and to select mutations efficiently. Here, we test the CRISPR technology in an emerging insect model and pest, the beetle Tribolium castaneum...
August 15, 2015: Development
https://www.readbyqxmd.com/read/25886725/transcriptome-differences-between-cry1ab-resistant-and-susceptible-strains-of-asian-corn-borer
#20
Li-Na Xu, Yue-Qin Wang, Zhen-Ying Wang, Ben-Jin Hu, Ying-Hui Ling, Kang-Lai He
BACKGROUND: Asian corn borer (ACB), Ostrinia furnacalis (Guenée), is the major insect pest of maize in China and countries of East and Southeast Asia, the Pacific and Australasia. ACB can develop strong resistance to the transgenic Bt maize expressing Cry1Ab, the most widely commercialized Bt maize worldwide. However, the molecular basis for the resistance mechanisms of ACB to Cry1Ab remained unclear. Two biological replicates of the transcriptome of Bt susceptible (ACB-BtS) and Cry1Ab resistant (ACB-AbR) strains of ACB were sequenced using Solexa/Illumina RNA-Seq technology to identify Cry1Ab resistance-relevant genes...
2015: BMC Genomics
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