caa a22 4500 605502994 CHVBK 20210128100608.0 cr unu---uuuuu 210128e20150901xx s 000 0 eng 10.1007/s00253-015-6755-2 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6755-2 Assembling of Holotrichia parallela (dark black chafer) midgut tissue transcriptome and identification of midgut proteins that bind to Cry8Ea toxin from Bacillus thuringiensis [Elektronische Daten] [Changlong Shu, Shuqian Tan, Jiao Yin, Mario Soberón, Alejandra Bravo, Chunqing Liu, Lili Geng, Fuping Song, Kebin Li, Jie Zhang] Holotrichia parallela is one of the most severe crop pests in China, affecting peanut, soybean, and sweet potato crops. Previous work showed that Cry8Ea toxin is highly effective against this insect. In order to identify Cry8Ea-binding proteins in the midgut cells of H. parallela larvae, we assembled a midgut tissue transcriptome by high-throughput sequencing and used this assembled transcriptome to identify Cry8Ea-binding proteins by liquid chromatography-tandem mass spectrometry (LC-MS/MS). First, we obtained de novo sequences of cDNAs from midgut tissue of H. parallela larvae and used available cDNA data in the GenBank. In a parallel assay, we obtained 11 Cry8Ea-binding proteins by pull-down assays performed with midgut brush border membrane vesicles. Peptide sequences from these proteins were matched to the H. parallela newly assembled midgut transcriptome, and 10 proteins were identified. Some of the proteins were shown to be intracellular proteins forming part of the cell cytoskeleton and/or vesicle transport such as actin, myosin, clathrin, dynein, and tubulin among others. In addition, an apolipophorin, which is a protein involved in lipid metabolism, and a novel membrane-bound alanyl aminopeptidase were identified. Our results suggest that Cry8Ea-binding proteins could be different from those characterized for Cry1A toxins in lepidopteran insects. Springer-Verlag Berlin Heidelberg, 2015 Bacillus thuringiensis nationallicence Cry8Ea nationallicence Binding nationallicence Holotrichia parallela nationallicence Midgut proteins nationallicence Shu Changlong State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut Tan Shuqian State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut Yin Jiao State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut Soberón Mario Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal 510-3, 62250, Cuernavaca, Morelos, Mexico aut Bravo Alejandra Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal 510-3, 62250, Cuernavaca, Morelos, Mexico aut Liu Chunqing Cangzhou Academy of Agricultural and Forestry Sciences, 061001, Cangzhou, People's Republic of China aut Geng Lili State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut Song Fuping State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut Li Kebin State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut Zhang Jie State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/17(2015-09-01), 7209-7218 0175-7598 99:17<7209 2015 99 253 https://doi.org/10.1007/s00253-015-6755-2 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6755-2 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Shu Changlong State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Tan Shuqian State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Yin Jiao State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Soberón Mario Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal 510-3, 62250, Cuernavaca, Morelos, Mexico aut NATIONALLICENCE 700 1- Bravo Alejandra Instituto de Biotecnología, Universidad Nacional Autónoma de México, Apdo. Postal 510-3, 62250, Cuernavaca, Morelos, Mexico aut NATIONALLICENCE 700 1- Liu Chunqing Cangzhou Academy of Agricultural and Forestry Sciences, 061001, Cangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Geng Lili State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Song Fuping State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Li Kebin State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Zhang Jie State Key Laboratory of Biology for Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, 100193, Beijing, People's Republic of China aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/17(2015-09-01), 7209-7218 0175-7598 99:17<7209 2015 99 253