Complete Genome Sequence of Bacillus thuringiensis Mutant Strain BMB171

Bacillus thuringiensis has been widely used as a biopesticide for a long time. Here we report the finished and annotated genome sequence of B. thuringiensis mutant strain BMB171, an acrystalliferous mutant strain with a high transformation frequency obtained and stocked in our laboratory.Bacillus thuringiensis is an insect pathogen which is widely used as a biopesticide due to its various endogenous crystal proteins and spores (12). To improve the virulence and practical effectiveness of B. thuringiensis, genetic transformation of different genes with beneficial traits is a fundamental procedure. Simultaneously, genetic transformation can facilitate functional genomic research. However, wild-type strains are not suitable to be used as recipient strains because of low transformation efficiency. This obstacle is mainly caused by the thick cell wall layer of B. thuringiensis together with multiple plasmids inside the cell, which harbor genes encoding insecticidal crystal proteins. We used the method of elevating the growth temperature and adding 0.05% sodium dodecyl sulfate to treat several parental strains and finally obtained mutant strain BMB171, with no resident plasmid, from wild-type crystalliferous strain YBT-1463 (9). The electrotransformation frequency of mutant BMB171 could reach up to 10⁷ transformants/μg DNA after optimization of the electrotransformation parameters (7), which was 4.8 × 10⁴-fold higher than that of the parental strain (8). Moreover, mutant strain BMB171 exhibited the same characteristics as YBT-1463, such as metabolic abilities and growth properties, as well as sensitivity to 10 antibiotics (8). Of course, BMB171 could produce parasporal crystals with characteristic geometric shapes through the expression of relevant cry genes carried by plasmids (7). Thus, B. thuringiensis mutant strain BMB171 has become a major recipient strain and is widely used for insecticidal crystal protein-encoding gene expression (14, 15), cell surface display (10, 13), gene function and regulation researches (2, 5), etc.The B. thuringiensis mutant strain BMB171 genome was sequenced by using a massive parallel pyrosequencing technology (454 GS-FLX). A total of 448,963 high-quality reads with an average read length of 391 bp were produced, providing about 32-fold coverage of the genome. Assembly was performed using the Newbler software of the 454 suite package (454 Life Sciences), which resulted in 193 large (defined as >500 bp) contigs. The relationship of contigs was determined by multiplex PCR, and gaps were filled through sequencing of PCR products by primer walking or shotgun sequencing with an ABI 3730 sequencer. The Phred/Phrap/Consed software package (3) was used for final sequence assembly and quality assessment. Protein-coding genes were predicted by combining the results of Glimmer 3.02 (1) and ZCURVE (4), followed by manual inspection. Both tRNA and rRNA genes were identified by tRNAscan-SE (11) and RNAmmer (6), respectively. Functional annotation was performed by searching against a protein database of the microbial genome developed in house.The 5.64-Mb genome of B. thuringiensis mutant strain BMB171 contains two replicons: a circular chromosome (5.33 Mb) encoding 5,088 open reading frames (ORFs) and a circular plasmid (0.31 Mb), which is named pBMB171, encoding 276 predicted ORFs. The G+C content of the chromosome is 35.3%, while that of the plasmid is 33.3%. The mutant strain BMB171 genome encodes 104 tRNAs and 14 rRNA operons. A previous study indicated that BMB171 is a plasmid-free mutant (9); however, our sequencing results demonstrated that a large plasmid still remains. The reason why the plasmid was not detected previously might be its large size and low copy number. We did not find any crystal protein genes in either chromosome or plasmid sequences, which was consistent with previous observations (9).In summary, the complete B. thuringiensis mutant strain BMB171 genome provides a better-defined genetic background for gene expression and regulation studies, especially crystal protein production and metabolic network construction.     

Complete Genome Sequence of Bacillus thuringiensis Serovar Sichuansis Strain MC28

Bacillus thuringiensis is an important microbial insecticide used in the control of agricultural pests. Here we report the finished, annotated genome sequence of Bacillus thuringiensis serovar Sichuansis strain MC28, which can form parasporal crystals consisting of Cry4Cc1, Cry30Fa1, Cry53Ab1, Cry54Aa1, Cry54Ab1, Cry68Aa1, Cry69Aa1, Cry69Aa2, Cry70Ba1, Cyt1Da1, and Cyt2Aa3. It is also highly toxic to lepidopterous and dipterous insects.     

Complete Genome Sequence of Mycoplasma hyorhinis Strain HUB-1

Mycoplasma hyorhinis is generally considered a swine pathogen yet is most commonly found infecting laboratory cell lines. An increasing body of evidence suggests that chronic infections with M. hyorhinis may cause oncogenic transformation. Here, we announce the complete genome sequence of M. hyorhinis strain HUB-1.Mycoplasma hyorhinis is generally considered to be a swine pathogen causing lung lesions, inflammation in the chest and abdominal lining, and arthritis (8). This agent also frequently contaminates laboratory cell cultures, impinging on many aspects of biological research (3). Recent studies have demonstrated that M. hyorhinis infections induce a malignant phenotype in human prostate (7) and gastric (4) cells, suggesting that M. hyorhinis infections are associated with oncogenic transformation. These properties of M. hyorhinis have increased its profile to researchers. The complete genome sequence of this microbe has yet to be determined.We sequenced the genome of M. hyorhinis strain HUB-1, a pathogenic strain isolated from the respiratory tract of swine. Whole-genome sequencing was performed by combining GS FLX (6) and Solexa paired-end sequencing technologies (1). Genomic libraries containing 3-kb inserts were constructed, and 308,604 reads (79.7% paired end) were produced using the GS FLX system, giving 65.9-fold coverage of the genome. About 93.4% of reads were assembled into one large scaffold using Newbler software (454 Life Sciences, Branford, CT). A total of 822,579 reads were generated using an Illumina Solexa Genome Analyzer IIx and were mapped to the scaffold using the Burrows-Wheeler alignment (BWA) tool (5). Gaps were filled by local assembly of the Solexa/Roche 454 reads or by sequencing PCR products by using an ABI 3730 capillary sequencer. Open reading frames containing more than 30 amino acid residues were predicted using Glimmer 3.0 (2) and verified by comparison with six other closely related genome sequences.The complete genome of M. hyorhinis HUB-1 consists of an 839,615-bp single circular chromosome with an average G+C content of 25.88%. A total of 654 protein-encoding genes are predicted. The average protein size is 364 amino acids, and the mean coding percentage is 85.2%. The genome includes 30 tRNA genes, and only a single copy of the 16S-23S rRNA operon can be found. The 5S rRNA operon is separate from the 16S-23S rRNA operon. Protein secretion occurs through a truncated membrane protein secretion system, consisting of SecA, SecD, SecY, PrsA, DnaK, Tig, and LepA. Additionally, 20 pseudogenes, which become truncated or inactivated, are identified in the genome.M. hyorhinis contains a special variable lipoprotein (Vlp) system that constitutes its major coat protein (9) and provides a mutational strategy for evasion of the host immune system. Different M. hyorhinis strains carry a variable number of vlp genes (9). M. hyorhinis HUB-1 is characterized to contain seven vlp genes displayed in the order 5′-vlpD-vlpE-vlpF-insertion sequence (IS)-vlpG-vlpA-IS-vlpB-vlpC-3′.This is the first complete genome sequence of M. hyorhinis, and its availability will provide a better-defined genetic background for future studies of gene expression and regulation.     

Complete Genome Sequence of the Diesel-Degrading Acinetobacter sp. Strain DR1

The genus Acinetobacter is ubiquitous in soil, aquatic, and sediment environments and includes pathogenic strains, such as A. baumannii. Many Acinetobacter species isolated from various environments have biotechnological potential since they are capable of degrading a variety of pollutants. Acinetobacter sp. strain DR1 has been identified as a diesel degrader. Here we report the complete genome sequence of Acinetobacter sp. DR1 isolated from the soil of a rice paddy.The genus Acinetobacter appears to be metabolically versatile and has the ability to degrade aliphatic hydrocarbon, thus making it an organism of interest for its possible bioremediational potential (9). Despite its biotechnological potential, the majority of genome projects conducted with Acinetobacter species have focused on pathogenic strains of A. baumannii. Currently, the only available whole-genome sequence of environmental isolates is that of A. baylyi ADP1 (2). Acinetobacter sp. strain DR1 was isolated from the soil of rice paddies, located in Deok-So (Korea University Agricultural Station), in the Kyonggi province of South Korea. Strain DR1 is capable of utilizing aliphatic hydrocarbons and diesel oil (5). Similarly to A. baylyi ADP1, this strain is also competent for natural transformation. We demonstrated previously that sodium chloride added to the medium induces the overproduction of exopolysaccharide (EPS), which evidences protective activity against diesel toxicity (4). Interestingly, DR1 possesses a quorum sensing (QS) system, which has been shown to play a significant role in biofilm formation and hexadecane biodegradation. The results of proteomic studies have demonstrated that the QS system regulates a broad variety of proteins (6). Collectively, our findings demonstrate that DR1 has profound potential for environmental applications and is an environmental isolate distinct from pathogenic strains, thus indicating that the whole-genome sequencing of DR1 is a worthwhile pursuit.Initial pyrosequencing using a GS-FLX system (454 Life Science Corporation) generated 652,162 reads (264,482,836 nucleotides; 64.3-fold coverage), which were assembled into 56 contigs. To determine the order of the contigs, 1,248 fosmid clones were constructed with an average insert size of 35 kb (10.5-fold coverage). The fosmid-end sequencing of 936 clones generated 1,372,452 bp. These high-quality Sanger reads allowed the assembly of 41 large contigs into 2 scaffolds containing 38 gaps. The gaps were filled via primer walking. All procedures for genome sequencing and gap filling were conducted by Macrogen (Seoul, South Korea). Protein coding regions were predicted with the GLIMMER3 software program (3), and automatic genome annotation was conducted on a RAST server (1) and the NCBI Prokaryotic Genomes Automatic Annotation Pipeline (PGAAP). The tRNA and rRNA genes were annotated using the tRNAScan-SE (8) and RNAmmer software programs (7), respectively. The genome of Acinetobacter sp. DR1 consists of a circular 4,152,543-bp chromosome with a G+C content of 38%, 3,874 predicted coding sequences, and 71 tRNAs. There are 6 rRNA operons with a 16S, tRNA-Ile, tRNA-Ala, 23S, 5S organization. The genes studied previously were clearly identified via genome sequencing (4, 5, 6). The availability of the complete genome sequence of Acinetobacter sp. strain DR1 will contribute to an in-depth understanding of the genetic potentials of Acinetobacter species.     

Complete Genome Sequence of Porcine Circovirus 2b Strain CC1

A porcine circovirus 2 (PCV2) strain, designated CC1, was isolated and purified from tissue samples from pigs with wasting syndromes in China. We report the complete genome sequence of PCV2b strain CC1 with a deletion of C at position 1053 resulting in elongation of open reading frame 2 (ORF2) and formation of ORF5. There were 11 ORFs in the genome.     

Complete Genome Sequence of Mycoplasma wenyonii Strain Massachusetts

Mycoplasma wenyonii is a hemotrophic mycoplasma that causes acute and chronic infections in cattle. Here, we announce the first complete genome sequence of this organism. The genome is a single circular chromosome with 650,228 bp and G+C% of 33.9. Analyses of M. wenyonii genome will provide insights into its biology.     

Complete Genome Sequence of Bacillus cereus NC7401, Which Produces High Levels of the Emetic Toxin Cereulide

We report the complete and annotated genome sequence of Bacillus cereus NC7401, a representative of the strain group that causes emetic-type food poisoning. The emetic toxin, cereulide, is produced by a nonribosomal protein synthesis (NRPS) system that is encoded by a gene cluster on a large resident plasmid, pNCcld.     

Complete Genome Sequence of Bacteriophage phiAC-1 Infecting Acinetobacter soli Strain KZ-1

To date, a number of bacteriophages (phages) infecting Acinetobacter species have been reported and characterized. However, Acinetobacter phages which infect A. soli have not been investigated yet. Here, we report the complete genome sequence of Acinetobacter phage phiAC-1, which belongs to the Myoviridae, infecting Acinetobacter soli strain KZ-1.     

Complete Genome Sequence of the Pyrene-Degrading Bacterium Cycloclasticus sp. Strain P1

Cycloclasticus sp. strain P1 was isolated from deep-sea sediments of the Pacific Ocean and characterized as a unique bacterium in the degradation of pyrene, a four-ring polycyclic aromatic hydrocarbon (PAH). Here we report the complete genome of P1 and genes associated with PAH degradation.     

Complete Genome Sequence of Streptococcus thermophilus Strain MN-ZLW-002

Streptococcus thermophilus MN-ZLW-002 was originally isolated from traditionally fermented Chinese dairy products. One of the strain-dependent characteristics of this bacterium is its ability to produce exopolysaccharides (EPSs). This study determined and analyzed the genome sequence of MN-ZLW-002. Its complete genome comprised 2,046 genes and 1,848,520 nucleotides with an average GC content of 39%. The EPS cluster of MN-ZLW-002 includes 25 open reading frames (ORFs), and some results indicate a horizontal gene transfer between MN-ZLW-002 and other lactic acid bacteria (LAB).     

Complete Sequence of the First Chimera Genome Constructed by Cloning the Whole Genome of Synechocystis Strain PCC6803 into the Bacillus subtilis 168 Genome

Genome synthesis of existing or designed genomes is made feasible by the first successful cloning of a cyanobacterium, Synechocystis PCC6803, in Gram-positive, endospore-forming Bacillus subtilis. Whole-genome sequence analysis of the isolate and parental B. subtilis strains provides clues for identifying single nucleotide polymorphisms (SNPs) in the 2 complete bacterial genomes in one cell.     

Complete Genome Sequence of the Naphthalene-Degrading Pseudomonas putida Strain ND6

Pseudomonas putida strain ND6 is an efficient naphthalene-degrading bacterium. The complete genome of strain ND6 was sequenced and annotated. The genes encoding the enzymes involved in catechol degradation by the ortho-cleavage pathway were found in the chromosomal sequence, which indicated that strain ND6 is able to metabolize naphthalene by the catechol meta- and ortho-cleavage pathways.     

Complete Genome Sequence of the Broad-Host-Range Strain Sinorhizobium fredii USDA257

Here we announce the complete genome sequence of the symbiotic and nitrogen-fixing bacterium Sinorhizobium fredii USDA257. The genome shares a high degree of sequence similarity with the closely related broad-host-range strains S. fredii NGR234 and HH103. Most strikingly, the USDA257 genome encodes a wealth of secretory systems.     

Complete Genome Sequence of Alcanivorax dieselolei Type Strain B5

Alcanivorax dieselolei B5^T was isolated from oil-contaminated surface water of the Bohai Sea of China and characterized by the efficient degradation of alkane (C₅-C₃₆). Here we report the complete genome of B5^T and genes associated with alkane degradation.     

Complete Genome Sequence of Mycobacterium vaccae Type Strain ATCC 25954

Mycobacterium vaccae is a rapidly growing, nontuberculous Mycobacterium species that is generally not considered a human pathogen and is of major pharmaceutical interest as an immunotherapeutic agent. We report here the annotated genome sequence of the M. vaccae type strain, ATCC 25954.     

Complete Genome Sequence of Porcine Circovirus 2b Strain Shandong

Shandong is a porcine circovirus 2b (PCV2b) strain that was isolated and purified from tissue samples from pigs with postweaning multisystemic wasting syndrome (PMWS) in the Shandong Province of China. Here, we report the complete genome sequence of strain Shandong, which may aid in understanding the molecular characteristics of this strain.     

Complete Genome Sequence of Porcine Circovirus 2d Strain GDYX

Porcine circovirus type 2 (PCV2) is the etiologic agent of porcine circovirus-associated disease, and it is mainly divided into five genotypes. Here, we report the complete genome sequence of PCV2 strain GDYX, which belongs to PCV2d and has a unique amino acid variation at position 169 (S to G).     

Complete Genome Sequence of Bacillus subtilis Strain QB928, a Strain Widely Used in B. subtilis Genetic Studies

The complete genome sequence of Bacillus subtilis strain QB928 was constructed to facilitate studies in the evolution of the genetic code. With a widespread use of the strain in Bacillus subtilis genetics studies, its complete genome sequence would facilitate deeper understanding of Bacillus subtilis genetics.     

Complete Genome Sequence of Klebsiella pneumoniae 1084, a Hypermucoviscosity-Negative K1 Clinical Strain

We report the complete genome sequence of Klebsiella pneumoniae 1084, a hypermucoviscosity-negative K1 clinical strain. Sequencing and annotation revealed a 5,386,705-bp circular chromosome (57.4% G+C content), which contains 4,962 protein-coding genes, 80 tRNA genes, and 25 rRNA genes.