The mitochondrial genome (mitogenome) is a vital tool in molecular phylogenetics, evolutionary biology, and population genetics. Here, we present a protocol for extracting mitochondrial DNA from shrimp (Macrobrachium rosenbergii) and prawn (Penaeus monodon) species using a differential centrifugation approach without relying on commercially available kits. We describe steps for sample preparation, crude mitochondrial extraction, mitochondrial DNA isolation, and purity assessment. We then detail procedures for confirming the presence of the mitogenome and sequencing the mitochondrial genome. 

Link: https://doi.org/10.1016/j.xpro.2025.103852 

We report the complete mitochondrial genome sequence of the giant freshwater prawn (Macrobrachium rosenbergii) from Bangladesh. The circular genome is 15,766 base pairs long, with a GC content of 38.1%, and contains 13 protein-coding genes, 22 tRNA genes, 2 rRNA genes, and 1 control region. 

Link: https://doi.org/10.1128/mra.00924-24

We report the complete mitochondrial genome sequence of the giant tiger prawn (Penaeus monodon) from Bangladesh. The circular genome spans 15,979 base pairs, with a GC content of 29.04%, and encodes 13 protein-coding genes, 22 tRNAs, 2 rRNAs, and 1 control region. 

Link: https://doi.org/10.1128/mra.00118-25 

Providencia rettgeri has increasingly been responsible for several infections, including urinary tract, post-burn wounds, neonatal sepsis, and others. The emergence of drug-resistant isolates of P rettgeri, accompanied by intrinsic and acquired antibiotic resistance, has exacerbated the challenge of treating such infections, necessitating the development of novel therapeutics. Hypothetical proteins (HPs) form a major portion of cellular proteins and can be targeted by these novel therapeutics. In this study, 410 HPs from a pan-drug-resistant (PDR) P rettgeri strain (MRSN845308) were functionally annotated and characterized by physicochemical properties, localization, virulence, essential ity, druggability, and functionality. Among 410 HPs, the VirulentPred 2.0 tool and VICMpred combinedly predicted 33 HPs as virulent, whereas 48 HPs were highly interacting proteins based on the STRING v12 database. BlastKOALA and eggNOG-mapper v2.1.12 predicted 13 HPs involved in several metabolic pathways like Riboflavin metabolism and Lipopolysaccharide biosynthesis. Overall, 83 HPs were selected as primary drug targets; however, only 80 remained after nonhomology searching and essentiality analysis. In addition, all were detected as novel drug targets according to DrugBank 5.1.12. Considering the potential of membrane and extracellular proteins, 29 HPs (extracellular, outer, and inner mem brane) were selected based on the combined prediction from PSORTb v3.0.3, CELLO v.2.5, BUSCA, SOSUIGramN, and PSLpred. According to the prevalence of those HPs in different strains of P rettgeri sequences in National Center for Biotechnology Information Identical Protein Groups (NCBI-IPG), 5 HPs were selected as final drug targets. In addition, 5 other HPs annotated as transporter proteins were also added to the list. As no crystal structures of our targets are present, 3-dimensional structures of selected HPs were predicted by the AlphaFold Server powered by AlphaFold 3. Our findings might facilitate a better understanding of the mechanism of virulence and pathogenesis, and up-to-date annotations can make uncharacterized HPs easy to identify as targets for novel therapeutics.