Skip to content
BY-NC-ND 3.0 license Open Access Published by De Gruyter Open Access November 25, 2015

Characterization of cry1Cb3 and cry1Fb7 from Bacillus thuringiensis subsp. galleriae

  • Tianpei Huang , Ying Xiao , Jieru Pan , Lingling Zhang , Ivan Gelbič and Xiong Guan
From the journal Open Life Sciences

Abstract

Two cry1-type genes encoding insecticidal crystal proteins (ICPs) were detected by PCR-RFLP and cloned from Bacillus thuringiensis subsp. galleriae 87. The nucleotide sequences were deposited in GenBank with accession numbers EU679501 and EU679502, and designated as cry1Fb7 and cry1Cb3 respectively by B. thuringiensis Delta- Endotoxin Nomenclature Committee. cry1Cb3 shared 99% homology with other cry1Cb genes. The existence of two additional stop codons indicated cry1Cb3 was a silent gene. The cry1Cb3 was 3531 bp with 38.98% G+C content and its first open reading frame (ORF) encoded a protein of 213 amino acid residues with a calculated molecular weight of 23.8 kDa and a predicted pI value of 4.63. Five amino acid sequence blocks (block 1, block 2, block 3, block 4 and block 5) were found in Cry1Cb3. Translation of cry1Fb7 revealed an ORF of 3525 bp with 39.12% G+C content and a protein with a calculated molecular weight of 133.2 kDa and a predicted pI value of 5.18. Cry1Fb7 had five amino acid sequence blocks (blocks 1, 2, 3, 4 and 5) and three domains (I, II and III), which consisted of 218 residues (Leu34 to Ala252), 197 residues (Thr257 to Asp454), and 138 residues (Ile464 to Glu600), respectively.

References

[1] Joung K.B., Cote J.C., Phylogenetic Analysis of Bacillus thuringiensis serovars based on 16S rRNA gene restriction fragments length polymorphism, J. Appl. Microbiol., 2001, 90, 115-122 10.1046/j.1365-2672.2001.01227.xSearch in Google Scholar PubMed

[2] Guan X., Progress in the studies and application of Bacillus thuringiensis, Review of China Agricultural Sciences and Technology, 2006, 8, 5-11 Search in Google Scholar

[3] Lecadet M.M., Frachon E., Cosmao Dumanoir V., Ripoteau H., Hamon S., Laurent P., et al., Updating the H-antigen classification of Bacillus thuringiensis, J. Appl. Microbiol., 1999. 86, 660-672. 10.1046/j.1365-2672.1999.00710.xSearch in Google Scholar PubMed

[4] Tsygankova S.V., Boulygina E.S., Ignatov A.N., Kuznetsov B.B., Korotkov E.V., Use of DIR-PCR for elaboration of molecular markers of intraspecies bacterial groups as exemplified by Bacillus thuringiensis, Mikrobiologiya, 77, 40-45 10.1134/S0026261708010050Search in Google Scholar

[5] Crickmore N., Zeigler D.R., Feitelson J., Schnepf E., Van Rie J., Lereclus D., et al., Revision of the nomenclature for the Bacillus thuringiensis pesticidal crystal proteins, Microbiol. Mol. Biol. Rev., 1998, 62, 807-813. 10.1128/MMBR.62.3.807-813.1998Search in Google Scholar PubMed PubMed Central

[6] Nariman A.H.A., PCR detection of cry genes in local Bacillus thuringiensis isolates, Aust..J. Basic Appl. Sci., 2007, 1, 461–466 Search in Google Scholar

[7] Tabashnik B.E., Dennehy Y.J., Sims M.A., Larkin K., Head G.P., Moa, W.J., et al., Control of resistant pink bollworm (Pectinophora gossypiella) by transgenic cotton that produces Bacillus thuringiensis toxin Cry2Ab, Appl. Environ. Microbiol., 2002, 68, 3790-3794 10.1128/AEM.68.8.3790-3794.2002Search in Google Scholar PubMed PubMed Central

[8] Tounsi S., Jaoua S., Characterization of a novel cry2Aa-type gene from Bacillus thuringiensis subsp. Kurstaki, Biotechnol. Lett., 2003, 25, 1219-1223 10.1023/A:1025016221891Search in Google Scholar

[9] Vilas-Bôas G.T., Lemos M.V., Diversity of cry genes and genetic characterization of Bacillus thuringiensis isolated from Brazil, Can. J. Microbiol., 2004, 50, 605–613. 10.1139/w04-052Search in Google Scholar PubMed

[10] Kuo W.S., Chak K.F., Identification of novel cry-type genes from Bacillus thuringiensis strains on the basis of restriction fragment length polymorphism of the PCR-amplified DNA, Appl. Environ. Microbiol., 1996, 62, 1369-1377 10.1128/aem.62.4.1369-1377.1996Search in Google Scholar PubMed PubMed Central

[11] Perani M., Bishop A.H., Vaid A., Prevalence of β-exotoxin, diarrhoeal toxin and specific δ-endotoxin in natural isolates of Bacillus thuringiensis, FEMS Microbiol. Lett., 1998, 160, 55-60 10.1111/j.1574-6968.1998.tb12890.xSearch in Google Scholar PubMed

[12] Song F.P., Zhang J., Huang D.F., Xie T.J., Zan Y.W., Dai L.Y., et al., Establishment of PCR-RFLP identification system of cry genes from Bacillus thuringiensis, Scientia Agricultuta Sinica, 1998, 31,13-18 Search in Google Scholar

[13] Lin Y., Huang Z.P., Chen J.W., Huang B.W., Guan X., Strategies and advances on PCR identification for insecticidal crystal protein genes of Bacillus thuringiensis, Chinese Journal of Agricultural Biotechnology, 2000, 8, 56-58 Search in Google Scholar

[14] Zhang J, Song F.P., Zuo Y.P., Dai L.Y., Huang D.F., Identification of cry-type genes of 31 Bacillus thuringiensis isolates and analysis of their expression product, Acta Microbiologica Sinica, 2000, 40, 372-378 Search in Google Scholar

[15] Lima A.S.G., Guidelli A.M., Abreu I.L., Lemos M.V.F., Identification of new isolates of Bacillus thuringiensis using rep-PCR products and δ-endotoxine electron microscopy, Genet. Mol. Biol., 2002, 25, 225-229. 10.1590/S1415-47572002000200017Search in Google Scholar

[16] Porcar M., Juárez-Pérez V., PCR-based identification of Bacillus thuringiensis pesticidal crystal genes, FEMS Microbiol. Rev., 2003, 26, 419-432 10.1111/j.1574-6976.2003.tb00624.xSearch in Google Scholar

[17] Wang J., Boets A., Van Rie J., Ren ,G., Characterization of cry1, cry2, and cry9 genes in Bacillus thuringiensis isolates from China, J. Invertebr. Pathol., 2003, 82, 63-71 10.1016/S0022-2011(02)00202-1Search in Google Scholar

[18] Yao J., Zhang J., Song F., Li, C., Liu, X., Huang D., Cloning and expression of cry1Ac gene from a novel Bacillus thuringiensis Ly30 strain, Chinese Journal of Agricultural Biotechnology, 2003, 11, 516-519 Search in Google Scholar

[19] Huang T.P., Characterization of important functional genes from Bacillus thuringiensis, Ph.D thesis, Fujian Agriculture and Forestry University, Fuzhou, China, 2005 Search in Google Scholar

[20] Chambers J.A., Jelen A., Gilbert M.P., Jany C.S., Johnson T.B., Gawron-Burke C., Isolation and characterization of a novel insecticidal crystal protein gene from Bacillus thuringiensis subsp aizawai, J. Bacteriol.,1991, 173, 3966-3976. 10.1128/jb.173.13.3966-3976.1991Search in Google Scholar PubMed PubMed Central

[21] Widner W.R,, Whiteley H.R., Two highly related insecticidal crystal proteins of Bacillus thuringiensis subsp. kurstaki possess different host range specificities, J. Bacteriol., 1989, 171, 965-974 10.1128/jb.171.2.965-974.1989Search in Google Scholar PubMed PubMed Central

[22] Stobdan T., Kaur S., Singh A., Cloning and nucleotide sequence of a novel cry gene from Bacillus thuringiensis. Biotechnol. Lett., 2004, 26, 1153-1156 10.1023/B:BILE.0000035488.85309.dfSearch in Google Scholar

[23] Kalman S., Kiehne K.L., Libs J.L., Tamamoto T., Cloning of a novel cry1C-type gene from a strain of Bacillus thuringiensis subsp. galleriae, Appl. Environ. Microbiol., 1993, 59, 1131-1137 10.1128/aem.59.4.1131-1137.1993Search in Google Scholar PubMed PubMed Central

[24] Visser B., Munsterman E., Stoker A., Wim G.D., A novel Bacillus thuringiensis gene encoding a Spodoptera exigua-specific crystal protein, J. Bacteriol., 1990, 172, 6783-6788 10.1128/jb.172.12.6783-6788.1990Search in Google Scholar PubMed PubMed Central

[25] Tan J., Zhang J., Song F., Chen Y., Wang, K., Huang D., The study on cloning and expression of B. thuringiensis cry1Ab13 gene. Acta Microbiologica Sinica, 2002, 42, 40-44 Search in Google Scholar

[26] Tang M., Tan L., Yu J., Zuan M., Pang, Y., Cloning and expression of ICP cry1Abl6 gene of Bacillus thuringiensis, Acta Microbiologica Sinica, 2003, 43, 417-423 Search in Google Scholar

[27] Huang Z., Guan C., Guan X., Cloning, characterization and expression of a new cry1Ab gene from Bacillus thuringiensis WB9, Biotechnol. Lett., 2004, 26, 1557-1561 10.1023/B:BILE.0000045652.00137.1fSearch in Google Scholar

[28] Naimov S., Weemen-Hendriks M., Dukiandjiev S., de Maaagd R.A., Bacillus thuringiensis delta-endotoxin Cry1 hybrid proteins with increased activity against the Colorado potato beetle, Appl. Environ. Microbiol., 2001, 67, 5328-5330 10.1128/AEM.67.11.5328-5330.2001Search in Google Scholar PubMed PubMed Central

[29] Sambrook J., Russell D.W., Molecular cloning - a laboratory manual, 3rd ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 2001 Search in Google Scholar

[30] Wu Z.L., Guo W.Z., Qiu J.Y., Huang T.P., Li X.B., Guan X., Cloning and localization of vip3A gene of Bacillus thuringiensis, Biotechnol. Lett., 2004, 26, 1425-1428 10.1023/B:BILE.0000045645.45536.3fSearch in Google Scholar

[31] Schnepf E., Crickmore N., Vanrie J.V., Ereclus D., Bacillus thuringiensis and its pesticidal crystal proteins, Microbiol. Mol. Biol. Rev., 1998, 62, 775-806 10.1128/MMBR.62.3.775-806.1998Search in Google Scholar PubMed PubMed Central

Received: 2015-2-12
Accepted: 2015-10-22
Published Online: 2015-11-25

©2015

This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Downloaded on 29.3.2024 from https://www.degruyter.com/document/doi/10.1515/biol-2015-0054/html
Scroll to top button