We found and characterized two cry8 genes from the Bacillus thuringiensis strain INTA Fr7-4 isolated in Argentina. These genes, cry8Kb3 and cry8Pa3, are located in a tandem array within a 13,200-bp DNA segment sequenced from a preparation of total DNA. They encode 1,169- and 1,176-amino-acid proteins, respectively. Both genes were cloned with their promoter sequences and the proteins were expressed separately in an acrystalliferous strain of B. thuringiensis leading to the formation of ovoid crystals in the recombinant strains. The toxicity against larvae of Anthonomus grandis Bh. (Coleoptera: Curculionidae) of a spore-crystal suspension from the recombinant strain containing cry8Pa3 was similar to that of the parent strain INTA Fr7-4.

Journal Section:
Research Article
Keywords:
Bacillus thuringiensis, cry8, Expression
1.
Amadio AF, Benintende GB, Zandomeni RO: Complete sequence of three plasmids from Bacillus thuringiensis INTA-FR7-4 environmental isolate and comparison with related plasmids from the Bacillus cereus group. Plasmid 2009;62:172-182.
[PubMed]
https://doi.org/10.1016/j.plasmid.2009.07.005
2.
Amadio AF, Navas LE, Sauka DH, Berretta MF, Benintende GB, Zandomeni RO: Identification, cloning and expression of an insecticide cry8 gene from Bacillus thuringiensis INTA Fr7-4. J Mol Microbiol Biotechnol 2013;23:401-409.
[PubMed]
https://doi.org/10.1159/000353206
3.
Asano S, Yamashita C, Iizuka T, Takeuchi K, Yamanaka S, Cerf D, Yamamoto T: A strain of Bacillus thuringiensis subsp. galleriae containing a novel cry8 gene highly toxic to Anomala cuprea (Coleoptera: Scarabaeidae). Biol Control 2003;28:191-196.
https://doi.org/10.1016/S1049-9644(03)00060-4
4.
Benintende G, Cozzi J: Actividad insecticida de distintas fracciones de cultivos líquidos de Bacillus moritai (Aizawa y Fujiyoshi) en Musca domestica L. (Diptera: Muscidae). RIA 1996;27:27-32.
5.
Bravo A, Gill SS, Soberón M: Mode of action of Bacillus thuringiensis Cry and Cyt toxins and their potential for insect control. Toxicon 2007;49:423-435.
[PubMed]
https://doi.org/10.1016/j.toxicon.2006.11.022
6.
Crickmore N, Baum J, Bravo A, Lereclus D, Narva K, Sampson K, Schnep E, Sun M, Zeigler DR: Bacillus thuringiensis toxin nomenclature. 2014. http://www.btnomenclature.info.
7.
Crickmore N, Ellar DJ: Involvement of a possible chaperonin in the efficient expression of a cloned CryIIA δ-endotoxin gene in Bacillus thuringiensis. Mol Microbiol 1992;6:1533-1537.
[PubMed]
https://doi.org/10.1111/j.1365-2958.1992.tb00874.x
8.
De Maagd RA, Bakker P, Staykov N, Dukiandjiev S, Stiekema W, Bosch D: Identification of Bacillus thuringiensis δ-endotoxin Cry1C domain III amino acid residues involved in insect specificity. Appl Environ Microbiol 1999;65:4369-4374.
[PubMed]
9.
Dervyn E, Poncet S, Klier A, Rapoport G: Transcriptional regulation of the cryIVD gene operon from Bacillus thuringiensis subsp. israelensis. J Bacteriol 1995;177:2283-2291.
[PubMed]
10.
Du L, Qiu L, Peng Q, Lereclus D, Zhang J, Song F, Huang D: Identification of the promoter in the intergenic region between orf1 and cry8Ea1 controlled by sigma H factor. Appl Environ Microbiol 2012;78:4164-4168.
[PubMed]
https://doi.org/10.1128/AEM.00622-12
11.
Herrero S, Gonzalez-Cabrera J, Ferre J, Bakker PL, de Maagd RA: Mutations in the Bacillus thuringiensis Cry1Ca toxin demonstrate the role of domains II and III in specificity towards Spodoptera exigua larvae. Biochem J 2004;384:507-513.
[PubMed]
https://doi.org/10.1042/BJ20041094
12.
Jones DT, Taylor WR, Thornton JM: The rapid generation of mutation data matrices from protein sequences. Comput Appl Biosci 1992;8:275-282.
[PubMed]
https://doi.org/10.1093/bioinformatics/8.3.275
13.
Jones LR, Zandomeni RO, Weber EL: A long distance RT-PCR able to amplify the Pestivirus genome. J Virol Methods 2006;134:197-204.
[PubMed]
https://doi.org/10.1016/j.jviromet.2006.01.005
14.
Kalendar R, Lee D, Schulman AH: FastPCR software for PCR primer and probe design and repeat search. Genes Genomes Genomics 2009;3:1-14.
15.
Larkin MA, Blackshields G, Brown NP, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG: Clustal W and Clustal X version 2.0. Bioinformatics 2007;23:2947-2948.
[PubMed]
https://doi.org/10.1093/bioinformatics/btm404
16.
Lereclus D, Arantes O, Chaufaux J, Lecadet M: Transformation and expression of a cloned δ-endotoxin gene in Bacillus thuringiensis. FEMS Microbiol Lett 1989;51:211-217.
[PubMed]
https://doi.org/10.1111/j.1574-6968.1989.tb03448.x
17.
Mahillon J, Lereclus D: Electroporation of Bacillus thuringiensis and Bacillus cereus; in Eynard N, Teissie J (eds): Electro-Transformation of Bacteria. Springer Lab Manual. Berlin, Springer, 2000, pp 242-252.
https://doi.org/10.1007/978-3-662-04305-9_30
18.
Mahillon J, Rezsöhazy R, Hallet B, Delcour J: IS231 and other Bacillus thuringiensis transposable elements: a review. Genetica 1994;93:13-26.
[PubMed]
https://doi.org/10.1007/BF01435236
19.
Nakasu EY, Firmino AA, Dias SC, Rocha TL, Ramos HB, Oliveira GR, Lucena W, Carlini CR, Grossi-de-Sá MF: Analysis of Cry8Ka5-binding proteins from Anthonomus grandis (Coleoptera: Curculionidae) midgut. J Invertebr Pathol 2010;104:227-230.
[PubMed]
https://doi.org/10.1016/j.jip.2010.01.012
20.
Ogiwara K, Hori H, Minami M, Takeuchi K, Sato R, Ohba M, Iwahana H: Nucleotide sequence of the gene encoding novel δ-endotoxin from Bacillus thuringiensis serovar japonensis strain Buibui specific to scarabaeid beetles. Curr Microbiol 1995;30:227-235.
[PubMed]
https://doi.org/10.1007/BF00293638
21.
Oliveira GR, Silva MC, Lucena WA, Nakasu EY, Firmino AA, Beneventi MA, Souza DS, Gomes JE Jr, de Souza JD Jr, Rigden DJ, Ramos HB, Soccol CR, Grossi-de-Sa MF: Improving Cry8Ka toxin activity towards the cotton boll weevil (Anthonomus grandis). BMC Biotechnol 2011;11:85.
[PubMed]
https://doi.org/10.1186/1472-6750-11-85
22.
Page RDM: TreeView: an application to display phylogenetic trees on personal computers. Comput Appl Biosci 1996;12:357-358.
[PubMed]
https://doi.org/10.1093/bioinformatics/12.4.357
23.
Roh JY, Jae YC, Ming SL, Byung RJ, Yeon HJ: Bacillus thuringiensis as a specific, safe, and effective tool for insect pest control. J Microbiol Biotechnol 2007;17:547-559.
[PubMed]
24.
Rutherford K, Parkhill J, Crook J, Horsnell T, Rice P, Rajandream MA, Barrell B: Artemis: sequence visualization and annotation. Bioinformatics 2000;16:944-945.
[PubMed]
https://doi.org/10.1093/bioinformatics/16.10.944
25.
Saitou N, Nei M: The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 1987;4:406-425.
[PubMed]
26.
Sambrook J, Fritsch E, Maniatis T: Molecular Cloning. A Laboratory Manual. Cold Spring Harbor, Cold Spring Harbor Laboratory Press, 1989.
https://doi.org/10.1016/0092-8674(90)90210-6
27.
Sauka D, Benintende G: Bacillus thuringiensis: general aspects. An approach to its use in the biological control of lepidopteran insects behaving as agricultural pests. Rev Argent Microbiol 2008;40:124-140.
[PubMed]
28.
Schneider-Orelli O: Entomologisches Praktikum, ed 2. Aarau, Sauerlander, 1947.
29.
Schnepf E, Crickmore N, Van Rie J, Lereclus D, Baum J, Feitelson J, Zeigler DR, Dean DH: Bacillus thuringiensis and its pesticidal crystal proteins. Microbiol Mol Biol Rev 1998;62:775-806.
[PubMed]
30.
Shu C, Yu H, Wang R, Fen S, Su X, Huang D, Zhang J, Song F: Characterization of two novel cry8 genes from Bacillus thuringiensis strain BT185. Curr Microbiol 2009;58:389-392.
[PubMed]
https://doi.org/10.1007/s00284-008-9338-y
31.
Staden R, Beal KF, Bonfield JK: The Staden package, 1998. Methods Mol Biol 1999;132:115-130.
[PubMed]
https://doi.org/10.1385/1-59259-192-2:115
32.
Widner WR, Whiteley HR: Two highly related insecticidal crystal proteins of Bacillus thuringiensis subsp. kurstaki possess different host range specificities. J Bacteriol 1989;171:965-974.
[PubMed]
33.
Wu D, Cao XL, Bay YY, Aronson AI: Sequence of an operon containing a novel δ-endotoxin gene from Bacillus thuringiensis. FEMS Microbiol Lett 1991;81:31-36.
[PubMed]
https://doi.org/10.1111/j.1574-6968.1991.tb04707.x
34.
Yu H, Zhang J, Huang D, Gao J, Song F: Characterization of Bacillus thuringiensis strain Bt185 toxic to the Asian cockchafer, Holotrichia parallela. Curr Microbiol 2006;53:13-17.
[PubMed]
https://doi.org/10.1007/s00284-005-0097-8
35.
Zhang J: Evolution by gene duplication: an update. Trends Ecol Evol 2003;18:292-298.
https://doi.org/10.1016/S0169-5347(03)00033-8
© 2014 S. Karger AG, Basel
Copyright / Drug Dosage / Disclaimer
Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher.
Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug.
Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.
2014
You do not currently have access to this content.