G Model
ARTICLE IN PRESS
BIOTEC 6971 1–2
Journal of Biotechnology xxx (2014) xxx–xxx
Contents lists available at ScienceDirect
Journal of Biotechnology journal homepage: www.elsevier.com/locate/jbiotec
Genome Announcement
1
Complete genome sequence of Bacillus thuringiensis serovar galleriae strain HD-29, a typical strain of commercial biopesticide
2
3
Q1
4 5
Lei Zhu, Jin-Shui Zheng, Qiu-Ling Gao, Yue-Ying Wang, Dong-Hai Peng, Li-Fang Ruan, Ming Sun ∗ State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei, People’s Republic of China
6 7 8
9 21
a r t i c l e
i n f o
a b s t r a c t
10 11 12 13 14
Article history: Received 17 December 2014 Accepted 22 December 2014 Available online xxx
15 16 17 18 19 20
Q2 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46
Keywords: Bacillus thuringiensis Complete genome Bt biopesticide Lepidoptera
Bacillus thuringiensis serovar galleriae is highly toxic to Lepidoptera insect pests, and has been widely used as Bt biopesticide in many countries. Here we reported the complete genome of strain HD-29, a standard serotype strain in galleriae serovariety. More than previous work reported, it harbors ten plasmids, and three large ones carry eight insecticidal protein genes (cry1Aa, cry1Ac, cry1Ca, cry1Da, cry1Ia, cry2Ab, cry9Ea and vip3Aa) and an intact zwittermicin A biosynthetic gene cluster. © 2014 Published by Elsevier B.V.
Bacillus thuringiensis produces insecticidal parasporal crystal proteins and is ubiquitous in natural environment (Schnepf et al., 1998; van Frankenhuyzen, 2009). Thus, tens of thousands of strains were isolated to find the most highly insecticidal toxic ones for Bt biopesticide commercialization, using various insect pest control fields (Roh et al., 2007). B. thuringiensis serovar galleriae (serotype H5a, 5b, Btg) strains are used as successful Bt biopesticide in many countries for a long time, such as “Entobacterin” in China and Russia (Chen and Fan, 1989; Kozlov et al., 1975; Shvetsova, 1962) and “Spicturin” in India (Shah, 1999). It particularly targeted to American Bollworm (Hellicoverpa armigera), Pink bollworm (Pectinophera species), spotted bollworm (Erias insulana), diamond back moth (Plutela xylostella (Linnaeus)) and other vegetable pests such as Colorado potato beetle (Leptinotarsa decemlineota (Say)) and forest insects (http://www.ncipm.org.in/bacillus-thuringiensisgalleria.htm). Herein, we present on the complete genome sequence of B. thuringiensis serovar galleriae strain NRRL HD-29, isolated from Dendrolimus sibericus in Czechoslovakia. It is a standard serotype strain in this species with a toxicity to Lepidoptera insect larvae (Fagundes et al., 2011). The HD-29 genome sequence will provide valuable information which will contribute to deeper understanding of entomopathogenicity in B. thuringiensis. The genome sequencing of B. thuringiensis strain HD-29 was performed on an Illumina HiSeq2500 instrument, using a 500 bp
∗ Corresponding author. Tel.: +86 27 87283455; fax: +86 27 87280670. E-mail address: [email protected] (M. Sun).
paired-end genomic library and 150 nucleotide (nt) length reads. The de novo assembly was performed by ABySS alignment tool version 1.3.7 (Simpson et al., 2009). Based on 8,160,035 pairs of the filtered reads, a total 264 contigs (>500 bp) were obtained by preliminarily assembly and the coverage is 389 folds. After using the completed B. thuringiensis genomes released in NCBI as references to arrange the contigs, most of the gaps between the contigs were rapidly closed using reads analysis and confirmed by PCR amplification. The genome annotation was performed by NCBI Prokaryotic Genome Automatic Annotation Pipeline (PGAAP, http://www.ncbi.nlm.nih.gov/books/NBK174280/). And the insecticidal protein genes were predicted by BtToxin scanner reported in our previous work (Ye et al., 2012). The total length of the HD-29 genome is 6,742,233 bp, with 34.9% average G+C content. Interestingly, the previous work showed that strain HD-29 harbors four plasmids (Fagundes et al., 2011), however, the assembly result showed that it includes a circular chromosome and ten plasmids: pBMB426 (426,282 bp), pBMB267 (267,359 bp), pBMB126 (126,898 bp), pBMB71 (71,373 bp), pBMB55 (55,460 bp), pBMB47 (46,979 bp), pBMBLin15 (14,749 bp), pBMB12 (12,866 bp), pBMB10 (10,656 bp) and pBMB8 (8423 bp) (Table 1). The G+C contents of these plasmids are from 29.6% to 39.9%, respectively. The annotation showed that the genome of strain HD-29 totally contains 6904 genes. The chromosome harbors 5890 genes, 42 rRNA and 114 tRNA genes (Table 1). Plasmid encodes 1014 genes, a tRNA gene and eight insecticidal protein genes. The comparative genomics analysis showed that large plasmid pBMB426 encodes two insecticidal parasporal protein genes
http://dx.doi.org/10.1016/j.jbiotec.2014.12.021 0168-1656/© 2014 Published by Elsevier B.V.
Please cite this article in press as: Zhu, L., et al., Complete genome sequence of Bacillus thuringiensis serovar galleriae strain HD-29, a typical strain of commercial biopesticide. J. Biotechnol. (2014), http://dx.doi.org/10.1016/j.jbiotec.2014.12.021
47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75
G Model
ARTICLE IN PRESS
BIOTEC 6971 1–2
L. Zhu et al. / Journal of Biotechnology xxx (2014) xxx–xxx
2 Table 1 Genome features of Bacillus thuringiensis HD-29. Features
Length (bp)
G+C content (%)
No. of genes
No. of tRNAs
No. of rRNAs
No. of insecticidal protein genes
Accession numbers
Chromosome Plasmid pBMB426 Plasmid pBMB267 Plasmid pBMB126 Plasmid pBMB71 Plasmid pBMB55 Plasmid pBMB47 Plasmid pBMBLin15 Plasmid pBMB12 Plasmid pBMB10 Plasmid pBMB8
5,701,188 426,282 267,359 126,898 71,373 55,460 46,979 14,749 12,866 10,656 8423
35.3 32.9 33.2 31.7 32.2 34.9 35.4 39.9 31.6 33.8 29.6
5890 354 244 139 85 57 68 24 20 13 10
114 1 – – – – – – – – –
42 – – – – – – – – – –
– 2 5 1 – – – – – – –
CP010089 CP010090 CP010091 CP010092 CP010093 CP010094 CP010095 CP010096 CP010097 CP010098 CP010099
Total
6,742,233
34.9
6904
115
42
8
–
95
(cry1Ca and cry1Da), and a intact zwittermicin A biosynthetic gene cluster, which was proved to have the ability to potentiate the insecticidal activity of the protein toxins produced by B. thuringiensis and a broad-spectrum antimicrobial activity (Broderick et al., 2000; Luo et al., 2011). Plasmid pBMB267 showed 99% homology with the large plasmid pCT281 in strain CT-43 (He et al., 2011) and encodes five insecticidal protein genes (cry1Aa, cry1Ia, cry2Ab, cry9Ea and vip3Aa). Plasmid pBMB126 encodes an insecticidal protein gene cry1Ac. And pBMB47 showed 99% homology with Bacillus phage Waukesha92 (Sauder et al., 2014). According to the insecticidal activity of B. thuringiensis crystal proteins (van Frankenhuyzen, 2009), besides the larvaes of Lepidoptera, the Cry proteins from HD-29 also are active to Coleoptera, Diptera (Cry1Aa, Cry1Ac, Cry1Ca, Cry1Da and Cry1Ia) and Hemiptera (Cry2Ab). All these toxins bring a wide insecticidal spectrum to strain HD-29. The availability of the HD-29 genome facilitates the understanding of toxin gene diversity and regulation, and is helpful for better usage and modification of this commercial strain in the insect pest control.
96
Nucleotide sequence accession number
76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94
103
This complete genome has been deposited at DDBJ/EMBL/ GenBank under the accession number CP010089–CP010099. The strain is available from the U.S. Department of Agriculture’s Agricultural Research Service (ARS) in Peoria under accession HD-29, Bacillus Genetic Stock Center (Columbus, USA) under accession 4G5, or Prof. Ming Sun (State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, PR China).
104
Acknowledgements
97 98 99 100 101 102
We thank Dr. D. Zeigler in BGSC provide the B. thuringiensis strain HD-29. This work was supported by grants from the 106 Q3 National High Technology Research and Development Program 107 (863) of China (2011AA10A203), China 948 Program of Ministry of 105
Agriculture (G25), the National Basic Research Program (973) of China (2009CB118902), the National Natural Science Foundation of China (31170047 and 31171901).
108 109 110
References Broderick, N.A., Goodman, R.M., Raffa, K.F., Handelsman, J., 2000. Synergy between zwittermicin A and Bacillus thuringiensis subsp. kurstaki against gypsy moth (Lepidoptera: Lymantriidae). Environ. Entomol. 29, 101–107. Chen, Q., Fan, Y.L., 1989. Molecular cloning and expression of Bacillus thuringiensis subsp. galleriae insecticidal crystal protein genes in Escherichia coli. Sci. China B 32, 830–836. Fagundes, R.B., Picoli, E.A., Lana, U.G., Valicente, F.H., 2011. Plasmid patterns of efficient and inefficient strains of Bacillus thuringiensis against Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae). Neotrop. Entomol. 40, 600–606. He, J., Wang, J., Yin, W., Shao, X., Zheng, H., Li, M., Zhao, Y., Sun, M., Wang, S., Yu, Z., 2011. Complete genome sequence of Bacillus thuringiensis subsp. chinensis strain CT-43. J. Bacteriol. 193, 3407–3408. Kozlov, M.P., Savel’ev, V.N., Reitblat, A.G., 1975. Effects of entobacterin on flea larvae. Med. Parazitol. (Mosk) 44, 89–92. Luo, Y., Ruan, L.F., Zhao, C.M., Wang, C.X., Peng, D.H., Sun, M., 2011. Validation of the intact zwittermicin A biosynthetic gene cluster and discovery of a complementary resistance mechanism in Bacillus thuringiensis. Antimicrob. Agents Chemother. 55, 4161–4169. Roh, J.Y., Choi, J.Y., Li, M.S., Jin, B.R., Je, Y.H., 2007. Bacillus thuringiensis as a specific, safe, and effective tool for insect pest control. J. Microbiol. Biotechnol. 17, 547–559. Sauder, A.B., Carter, B., Langouet Astrie, C., Temple, L., 2014. Complete genome sequence of a mosaic bacteriophage, waukesha92. Genome Announc. 2, e0033900314. Schnepf, E., Crickmore, N., Van Rie, J., Lereclus, D., Baum, J., Feitelson, J., Zeigler, D.R., Dean, D.H., 1998. Bacillus thuringiensis and its pesticidal crystal proteins. Microbiol. Mol. Biol. Rev. 62, 775–806. Shah, P.A., 1999. Directory of Microbial Control Products and Services. http://www.sipweb.org/docs/mcddir.pdf Shvetsova, O.I., 1962. Biological basis of the effectiveness of the bacterial preparation entobacterin-3 for control of harmful insects. Zhurnal obshchei biologii 23, 381–390. Simpson, J.T., Wong, K., Jackman, S.D., Schein, J.E., Jones, S.J., Birol, I., 2009. ABySS: a parallel assembler for short read sequence data. Genome Res. 19, 1117–1123. van Frankenhuyzen, K., 2009. Insecticidal activity of Bacillus thuringiensis crystal proteins. J. Invertebr. Pathol. 101, 1–16. Ye, W., Zhu, L., Liu, Y., Crickmore, N., Peng, D., Ruan, L., Sun, M., 2012. Mining new crystal protein genes from Bacillus thuringiensis on the basis of mixed plasmid-enriched genome sequencing and a computational pipeline. Appl. Environ. Microbiol. 78, 4795–4801.
Please cite this article in press as: Zhu, L., et al., Complete genome sequence of Bacillus thuringiensis serovar galleriae strain HD-29, a typical strain of commercial biopesticide. J. Biotechnol. (2014), http://dx.doi.org/10.1016/j.jbiotec.2014.12.021
111
112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151
ARTICLE IN PRESS
BIOTEC 6971 1–2
Journal of Biotechnology xxx (2014) xxx–xxx
Contents lists available at ScienceDirect
Journal of Biotechnology journal homepage: www.elsevier.com/locate/jbiotec
Genome Announcement
1
Complete genome sequence of Bacillus thuringiensis serovar galleriae strain HD-29, a typical strain of commercial biopesticide
2
3
Q1
4 5
Lei Zhu, Jin-Shui Zheng, Qiu-Ling Gao, Yue-Ying Wang, Dong-Hai Peng, Li-Fang Ruan, Ming Sun ∗ State Key Laboratory of Agricultural Microbiology, College of Life Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei, People’s Republic of China
6 7 8
9 21
a r t i c l e
i n f o
a b s t r a c t
10 11 12 13 14
Article history: Received 17 December 2014 Accepted 22 December 2014 Available online xxx
15 16 17 18 19 20
Q2 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46
Keywords: Bacillus thuringiensis Complete genome Bt biopesticide Lepidoptera
Bacillus thuringiensis serovar galleriae is highly toxic to Lepidoptera insect pests, and has been widely used as Bt biopesticide in many countries. Here we reported the complete genome of strain HD-29, a standard serotype strain in galleriae serovariety. More than previous work reported, it harbors ten plasmids, and three large ones carry eight insecticidal protein genes (cry1Aa, cry1Ac, cry1Ca, cry1Da, cry1Ia, cry2Ab, cry9Ea and vip3Aa) and an intact zwittermicin A biosynthetic gene cluster. © 2014 Published by Elsevier B.V.
Bacillus thuringiensis produces insecticidal parasporal crystal proteins and is ubiquitous in natural environment (Schnepf et al., 1998; van Frankenhuyzen, 2009). Thus, tens of thousands of strains were isolated to find the most highly insecticidal toxic ones for Bt biopesticide commercialization, using various insect pest control fields (Roh et al., 2007). B. thuringiensis serovar galleriae (serotype H5a, 5b, Btg) strains are used as successful Bt biopesticide in many countries for a long time, such as “Entobacterin” in China and Russia (Chen and Fan, 1989; Kozlov et al., 1975; Shvetsova, 1962) and “Spicturin” in India (Shah, 1999). It particularly targeted to American Bollworm (Hellicoverpa armigera), Pink bollworm (Pectinophera species), spotted bollworm (Erias insulana), diamond back moth (Plutela xylostella (Linnaeus)) and other vegetable pests such as Colorado potato beetle (Leptinotarsa decemlineota (Say)) and forest insects (http://www.ncipm.org.in/bacillus-thuringiensisgalleria.htm). Herein, we present on the complete genome sequence of B. thuringiensis serovar galleriae strain NRRL HD-29, isolated from Dendrolimus sibericus in Czechoslovakia. It is a standard serotype strain in this species with a toxicity to Lepidoptera insect larvae (Fagundes et al., 2011). The HD-29 genome sequence will provide valuable information which will contribute to deeper understanding of entomopathogenicity in B. thuringiensis. The genome sequencing of B. thuringiensis strain HD-29 was performed on an Illumina HiSeq2500 instrument, using a 500 bp
∗ Corresponding author. Tel.: +86 27 87283455; fax: +86 27 87280670. E-mail address: [email protected] (M. Sun).
paired-end genomic library and 150 nucleotide (nt) length reads. The de novo assembly was performed by ABySS alignment tool version 1.3.7 (Simpson et al., 2009). Based on 8,160,035 pairs of the filtered reads, a total 264 contigs (>500 bp) were obtained by preliminarily assembly and the coverage is 389 folds. After using the completed B. thuringiensis genomes released in NCBI as references to arrange the contigs, most of the gaps between the contigs were rapidly closed using reads analysis and confirmed by PCR amplification. The genome annotation was performed by NCBI Prokaryotic Genome Automatic Annotation Pipeline (PGAAP, http://www.ncbi.nlm.nih.gov/books/NBK174280/). And the insecticidal protein genes were predicted by BtToxin scanner reported in our previous work (Ye et al., 2012). The total length of the HD-29 genome is 6,742,233 bp, with 34.9% average G+C content. Interestingly, the previous work showed that strain HD-29 harbors four plasmids (Fagundes et al., 2011), however, the assembly result showed that it includes a circular chromosome and ten plasmids: pBMB426 (426,282 bp), pBMB267 (267,359 bp), pBMB126 (126,898 bp), pBMB71 (71,373 bp), pBMB55 (55,460 bp), pBMB47 (46,979 bp), pBMBLin15 (14,749 bp), pBMB12 (12,866 bp), pBMB10 (10,656 bp) and pBMB8 (8423 bp) (Table 1). The G+C contents of these plasmids are from 29.6% to 39.9%, respectively. The annotation showed that the genome of strain HD-29 totally contains 6904 genes. The chromosome harbors 5890 genes, 42 rRNA and 114 tRNA genes (Table 1). Plasmid encodes 1014 genes, a tRNA gene and eight insecticidal protein genes. The comparative genomics analysis showed that large plasmid pBMB426 encodes two insecticidal parasporal protein genes
http://dx.doi.org/10.1016/j.jbiotec.2014.12.021 0168-1656/© 2014 Published by Elsevier B.V.
Please cite this article in press as: Zhu, L., et al., Complete genome sequence of Bacillus thuringiensis serovar galleriae strain HD-29, a typical strain of commercial biopesticide. J. Biotechnol. (2014), http://dx.doi.org/10.1016/j.jbiotec.2014.12.021
47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75
G Model
ARTICLE IN PRESS
BIOTEC 6971 1–2
L. Zhu et al. / Journal of Biotechnology xxx (2014) xxx–xxx
2 Table 1 Genome features of Bacillus thuringiensis HD-29. Features
Length (bp)
G+C content (%)
No. of genes
No. of tRNAs
No. of rRNAs
No. of insecticidal protein genes
Accession numbers
Chromosome Plasmid pBMB426 Plasmid pBMB267 Plasmid pBMB126 Plasmid pBMB71 Plasmid pBMB55 Plasmid pBMB47 Plasmid pBMBLin15 Plasmid pBMB12 Plasmid pBMB10 Plasmid pBMB8
5,701,188 426,282 267,359 126,898 71,373 55,460 46,979 14,749 12,866 10,656 8423
35.3 32.9 33.2 31.7 32.2 34.9 35.4 39.9 31.6 33.8 29.6
5890 354 244 139 85 57 68 24 20 13 10
114 1 – – – – – – – – –
42 – – – – – – – – – –
– 2 5 1 – – – – – – –
CP010089 CP010090 CP010091 CP010092 CP010093 CP010094 CP010095 CP010096 CP010097 CP010098 CP010099
Total
6,742,233
34.9
6904
115
42
8
–
95
(cry1Ca and cry1Da), and a intact zwittermicin A biosynthetic gene cluster, which was proved to have the ability to potentiate the insecticidal activity of the protein toxins produced by B. thuringiensis and a broad-spectrum antimicrobial activity (Broderick et al., 2000; Luo et al., 2011). Plasmid pBMB267 showed 99% homology with the large plasmid pCT281 in strain CT-43 (He et al., 2011) and encodes five insecticidal protein genes (cry1Aa, cry1Ia, cry2Ab, cry9Ea and vip3Aa). Plasmid pBMB126 encodes an insecticidal protein gene cry1Ac. And pBMB47 showed 99% homology with Bacillus phage Waukesha92 (Sauder et al., 2014). According to the insecticidal activity of B. thuringiensis crystal proteins (van Frankenhuyzen, 2009), besides the larvaes of Lepidoptera, the Cry proteins from HD-29 also are active to Coleoptera, Diptera (Cry1Aa, Cry1Ac, Cry1Ca, Cry1Da and Cry1Ia) and Hemiptera (Cry2Ab). All these toxins bring a wide insecticidal spectrum to strain HD-29. The availability of the HD-29 genome facilitates the understanding of toxin gene diversity and regulation, and is helpful for better usage and modification of this commercial strain in the insect pest control.
96
Nucleotide sequence accession number
76 77 78 79 80 81 82 83 84 85 86 87 88 89 90 91 92 93 94
103
This complete genome has been deposited at DDBJ/EMBL/ GenBank under the accession number CP010089–CP010099. The strain is available from the U.S. Department of Agriculture’s Agricultural Research Service (ARS) in Peoria under accession HD-29, Bacillus Genetic Stock Center (Columbus, USA) under accession 4G5, or Prof. Ming Sun (State Key Laboratory of Agricultural Microbiology, Huazhong Agricultural University, Wuhan, PR China).
104
Acknowledgements
97 98 99 100 101 102
We thank Dr. D. Zeigler in BGSC provide the B. thuringiensis strain HD-29. This work was supported by grants from the 106 Q3 National High Technology Research and Development Program 107 (863) of China (2011AA10A203), China 948 Program of Ministry of 105
Agriculture (G25), the National Basic Research Program (973) of China (2009CB118902), the National Natural Science Foundation of China (31170047 and 31171901).
108 109 110
References Broderick, N.A., Goodman, R.M., Raffa, K.F., Handelsman, J., 2000. Synergy between zwittermicin A and Bacillus thuringiensis subsp. kurstaki against gypsy moth (Lepidoptera: Lymantriidae). Environ. Entomol. 29, 101–107. Chen, Q., Fan, Y.L., 1989. Molecular cloning and expression of Bacillus thuringiensis subsp. galleriae insecticidal crystal protein genes in Escherichia coli. Sci. China B 32, 830–836. Fagundes, R.B., Picoli, E.A., Lana, U.G., Valicente, F.H., 2011. Plasmid patterns of efficient and inefficient strains of Bacillus thuringiensis against Spodoptera frugiperda (J.E. Smith) (Lepidoptera: Noctuidae). Neotrop. Entomol. 40, 600–606. He, J., Wang, J., Yin, W., Shao, X., Zheng, H., Li, M., Zhao, Y., Sun, M., Wang, S., Yu, Z., 2011. Complete genome sequence of Bacillus thuringiensis subsp. chinensis strain CT-43. J. Bacteriol. 193, 3407–3408. Kozlov, M.P., Savel’ev, V.N., Reitblat, A.G., 1975. Effects of entobacterin on flea larvae. Med. Parazitol. (Mosk) 44, 89–92. Luo, Y., Ruan, L.F., Zhao, C.M., Wang, C.X., Peng, D.H., Sun, M., 2011. Validation of the intact zwittermicin A biosynthetic gene cluster and discovery of a complementary resistance mechanism in Bacillus thuringiensis. Antimicrob. Agents Chemother. 55, 4161–4169. Roh, J.Y., Choi, J.Y., Li, M.S., Jin, B.R., Je, Y.H., 2007. Bacillus thuringiensis as a specific, safe, and effective tool for insect pest control. J. Microbiol. Biotechnol. 17, 547–559. Sauder, A.B., Carter, B., Langouet Astrie, C., Temple, L., 2014. Complete genome sequence of a mosaic bacteriophage, waukesha92. Genome Announc. 2, e0033900314. Schnepf, E., Crickmore, N., Van Rie, J., Lereclus, D., Baum, J., Feitelson, J., Zeigler, D.R., Dean, D.H., 1998. Bacillus thuringiensis and its pesticidal crystal proteins. Microbiol. Mol. Biol. Rev. 62, 775–806. Shah, P.A., 1999. Directory of Microbial Control Products and Services. http://www.sipweb.org/docs/mcddir.pdf Shvetsova, O.I., 1962. Biological basis of the effectiveness of the bacterial preparation entobacterin-3 for control of harmful insects. Zhurnal obshchei biologii 23, 381–390. Simpson, J.T., Wong, K., Jackman, S.D., Schein, J.E., Jones, S.J., Birol, I., 2009. ABySS: a parallel assembler for short read sequence data. Genome Res. 19, 1117–1123. van Frankenhuyzen, K., 2009. Insecticidal activity of Bacillus thuringiensis crystal proteins. J. Invertebr. Pathol. 101, 1–16. Ye, W., Zhu, L., Liu, Y., Crickmore, N., Peng, D., Ruan, L., Sun, M., 2012. Mining new crystal protein genes from Bacillus thuringiensis on the basis of mixed plasmid-enriched genome sequencing and a computational pipeline. Appl. Environ. Microbiol. 78, 4795–4801.
Please cite this article in press as: Zhu, L., et al., Complete genome sequence of Bacillus thuringiensis serovar galleriae strain HD-29, a typical strain of commercial biopesticide. J. Biotechnol. (2014), http://dx.doi.org/10.1016/j.jbiotec.2014.12.021
111
112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139 140 141 142 143 144 145 146 147 148 149 150 151
