Genome assembly and comparative genomic analysis dataset of Bacillus thuringiensis strain BT-MFS
Description
This dataset presents the comprehensive whole-genome sequencing (WGS) data and extensive bioinformatics pipeline outputs for *Bacillus thuringiensis* strain BT-MFS, an isolate recovered from a dead Moroccan locust (*Dociostaurus maroccanus*) collected in Boysun, Surkhandarya, Uzbekistan. Genomic DNA was sequenced on the DNBSEQ platform utilizing a paired-end 150-bp strategy. Following initial read quality assessment with FastQC, *de novo* genome assembly was executed using SPAdes, and the resulting assembly's contiguity and completeness were rigorously evaluated via QUAST, BUSCO, and CheckM2. High-resolution taxonomic classification and evolutionary positioning were established using Kraken2, GTDB-Tk, the Type (Strain) Genome Server (TYGS), and Average Nucleotide Identity (FastANI) calculations. Structural genome annotation, including the specific identification of insecticidal *Cry* genes, was performed using Prokka. This was significantly expanded with deep functional profiling and secondary metabolite biosynthetic gene cluster (BGC) predictions utilizing eggNOG-mapper, InterProScan, and antiSMASH. Furthermore, the dataset encompasses detailed characterizations of the mobilome and bacterial defense mechanisms, providing detection and reconstruction outputs for plasmids (PlasmidFinder, MOB-suite), prophages (Phastest), and CRISPR arrays. Finally, core-genome phylogenetic and comparative pangenomic analyses against related *Bacillus* reference strains were conducted using Panaroo and IQ-TREE, complemented by circular genome visualizations generated via Proksee. For full reproducibility, all intermediate files, final outputs, and an organized file inventory are included in the repository.
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Steps to reproduce
The BT-MFS strain was isolated from a dead Dociostaurus maroccanus grasshopper collected in Boysun, Surkhandarya, Uzbekistan. Genomic DNA was extracted using a modified Marmur protocol [1], followed by whole-genome sequencing on the DNBSEQ platform using paired-end 150-bp reads with an approximately 350-bp insert size. Raw reads were assessed using FastQC and processed for de novo genome assembly using SPAdes. Contigs shorter than 1000 bp were excluded from the final assembly. Assembly quality was evaluated using QUAST, BUSCO, and CheckM2. Taxonomic classification was performed using Kraken2 and GTDB-Tk. Structural genome annotation was performed using Prokka, followed by functional annotation and protein characterization using eggNOG-mapper and InterProScan. Secondary metabolite biosynthetic gene clusters were investigated using antiSMASH. CRISPR-Cas elements were identified using CRISPR-focused genome analysis tools, and prophage sequences were assessed using PHASTEST. Comparative genomic analysis was performed using FastANI, while core-genome analysis was conducted using Panaroo followed by maximum-likelihood phylogenetic reconstruction using IQ-TREE 3. Genome-based taxonomic analysis was additionally performed using TYGS. The resulting sequencing reads, genome assembly, quality-control reports, annotation outputs, functional and secondary-metabolite analyses, CRISPR-Cas and prophage results, taxonomic analyses, and phylogenetic datasets were compiled for data deposition and reuse.