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Results for “molecular biology”

Search indexed bioRxiv preprints in genomics, neuroscience, cell biology and bioinformatics. Read source abstracts and check manuscript versions; preprints are not peer reviewed.

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At least 127 records · Page 7Linked to original sources

bioRxiv2026

T cell-derived IFNγ instructs ECM crosslinking by cardiac fibroblasts through LOXL3 in experimental cardiometabolic HFpEF

BackgroundHeart failure with preserved ejection fraction (HFpEF) is a major clinical challenge characterized by diastolic dysfunction. Left ventricular stiffening and inflammation are hallmarks of HFpEF, yet the contribution of extracellular matrix (ECM) stiffness and the immune-stromal mechanisms driving ECM stiffening in cardiometabolic HFpEF remain poorly understood. MethodsWe used the murine "2-hit model" of cardiometabolic HFpEF, in which the combination of high fat diet and hypertension induced by L-NAME causes diastolic dysfunction. We evaluated diastolic function by echocardiography and ECM mechanics by uniaxial tensile testing of decellularized cardiac tissue. Functional in vivo studies included genetic depletion of T cells, interferon-{gamma} (IFN{gamma}) knockout mice, and pharmacological lysyl oxidase inhibition. We combined co-cultures of CD4+ T cells and cardiac fibroblasts (CFB) with mechanical testing of cardiac ECM and molecular biology to elucidate cellular and molecular mechanisms. ResultsLeft ventricular ECM stiffness strongly correlated with impaired diastolic function in experimental cardiometabolic HFpEF. Cardiac CD4 T cell infiltration was required for ECM stiffening and upregulation of lysyl oxidase enzymes in CFB. CD4+ T cell-derived IFN{gamma} was both necessary and sufficient to induce LOXL3 in CFB, which increased ECM stiffness in vitro. Mechanistically, IFN{gamma} signaling activated hypoxia-inducible factor-1 (HIF1) in CFB, driving LOXL3 expression and subsequent collagen crosslinking. Genetic or pharmacologic disruption of this IFN{gamma}-HIF1-LOXL3 axis in vivo attenuated adverse ECM remodeling and improved diastolic function. ConclusionsCD4 T cells promote pathological ECM stiffening in cardiometabolic HFpEF through IFN{gamma}-mediated, LOXL3-dependent ECM crosslinking by CFB. Targeting this immune-stromal pathway may offer a novel therapeutic strategy for HFpEF.

pathology↗

bioRxiv2025

SuB3 a Simple Subcellular Fractionation Protocol for Localization Studies of Nucleic Acids and Proteins

Understanding the subcellular localization of RNA and proteins is critical to dissecting gene regulation in eukaryotic organisms. However, this task is elusive as existing fractionation methods often rely on protoplast isolation or commercial kits, that are labor-intensive, costly, and can introduce stress-induced transcriptomic and proteomic changes. Here, we present a simple, rapid, and cost-effective protocol for the fractionation of nuclear and cytoplasmic components directly from diverse plant tissues, that does not require protoplastization. This Subcellular-fractionation protocol in 3 steps (a.k.a. "bueno, bonito y barato"- spanish for "good, nice and cheap"-), referred to as "SuB3", yields nuclear- and cytoplasmic- enriched subcellular fractions suitable for downstream applications such as RT-PCR, RNA/cDNA sequencing, and Western blotting. The procedure is based on sequential detergent-assisted extraction and centrifugation and enables the simultaneous isolation of RNA and protein from the same biological material. Due to its simplicity, speed, and broad compatibility, this protocol is a valuable tool for plant molecular biology laboratories investigating subcellular dynamics of gene expression.

molecular biology↗

bioRxiv2026

CRISPR/Cas9 gene editing in the heterotrophic protist Acanthamoeba castellanii

Members of the amoebozoan genus Acanthamoeba are unicellular heterotrophic protists that inhabit a wide range of terrestrial and aquatic environments. They are opportunistic pathogens known to host disease-causing bacteria as well large DNA viruses. While Acanthamoeba is commonly used to study cellular locomotion, phagocytosis, and disease, genetic tools for precise editing of the Acanthamoeba genome are still in their infancy. Here we have developed CRISPR/Cas9 tools for genetically engineering the model acanthamoebid Acanthamoeba castellanii strain Neff. In proof-of-principle experiments, we targeted the myosin-II heavy chain gene. Evidence for editing was found using reverse transcriptase PCR, microscopy, and DNA sequencing, which also identified unique alleles for the target locus. Collectively, we demonstrate that CRISPR can be employed to edit Acanthamoeba genes using a knock-in approach. This work serves as a foundation to further develop A. castellanii as a model system with which to study diverse questions in cell and molecular biology, biochemistry and evolution.

molecular biology↗

bioRxiv2015

Biological screens from linear codes: theory and tools

Molecular biology increasingly relies on large screens where enormous numbers of specimens are systematically assayed in the search for a particular, rare outcome. These screens include the systematic testing of small molecules for potential drugs and testing the association between genetic variation and a phenotype of interest. While these screens are \"hypothesis-free,\" they can be wasteful; pooling the specimens and then testing the pools is more efficient. We articulate in precise mathematical ways the type of structures useful in combinatorial pooling designs so as to eliminate waste, to provide light weight, flexible, and modular designs. We show that Reed-Solomon codes, and more generally linear codes, satisfy all of these mathematical properties. We further demonstrate the power of this technique with Reed-Solomonbased biological experiments. We provide general purpose tools for experimentalists to construct and carry out practical pooling designs with rigorous guarantees for large screens.

Bioinformatics↗

bioRxiv2018

Golden Mutagenesis: An efficient multi-site saturation mutagenesis approach by Golden Gate cloning with automated primer design

Site-directed methods for the generation of genetic diversity are essential tools in the field of directed enzyme evolution. The Golden Gate cloning technique has been proven to be an efficient tool for a variety of cloning setups. The utilization of restriction enzymes which cut outside of their recognition domain allows the assembly of multiple gene fragments obtained by PCR amplification without altering the open reading frame of the reconstituted gene. We have developed a protocol, termed Golden Muta-genesis that allows the rapid, straightforward, reliable and inexpensive construction of mutagenesis libraries. One to five amino acid positions within a coding sequence could be altered simultaneously using a protocol which can be performed within one day. To facilitate the implementation of this technique, a software library and web application for automated primer design and for the graphical evaluation of the randomization success based on the sequencing results was developed. This allows facile primer design and application of Golden Mutagenesis also for laboratories, which are not specialized in molecular biology.

molecular biology↗

bioRxiv2022

During replicative aging, changes in DNA double-strand break repair correlate with an increased rate of mutation.

A double -strand break (DSB) is one of the most deleterious forms of DNA damage. In eukaryotic cells, two main repair pathways have evolved to repair DSBs, homologous recombination (HR) and non-homologous end-joining (NHEJ). HR is the predominant pathway of repair in the unicellular eukaryotic organism, S. cerevisiae. However, during replicative aging the relative use of HR and NHEJ shifts in favor of end-joining repair. By monitoring repair events in the HO-DSB system, we find that early in replicative aging there is a decrease in the association of long-range resection factors, Dna2-Sgs1 and Exo1 at the break site and a decrease in DNA resection. Subsequently, as aging progressed, the recovery of Ku70 at DSBs decreased and the break site associated with the nuclear pore complex at the nuclear periphery, which is the location where DSB repair occurs through alternative pathways that are more mutagenic. End-bridging remained intact as HR and NHEJ declined, but eventually it too became disrupted in cells at advanced replicative age. In all, our work provides insight into the molecular changes in DSB repair pathway during replicative aging. HR first declined, resulting in a transient increase in the NHEJ. However, with increased cellular divisions, Ku70 recovery at DSBs and NHEJ subsequently declined. In wild type cells of advanced replicative age, there was a high frequency of repair products with genomic deletions and microhomologies at the break junction, events not observed in young cells which repaired primarily by HR. HighlightsO_LIDecreased DNA resection at DSBs is an early event of replicative aging C_LIO_LIEnd-joining repair increases as resection decreases at DSBs in older cells C_LIO_LIIn older cells the products of DSB repair contain deletions and microhomologies C_LIO_LIDSBs associate with the NPC at the nuclear periphery more in older cells C_LIO_LIOld Cell Enrichment method suitable for molecular biology approaches in budding yeast C_LI

molecular biology↗

bioRxiv2024

Leaving Science: Attrition of Biologists in 38 OECD Countries

This study examines biologists leaving science in 38 OECD countries in the past two decades. We use publication metadata from a global bibliometric database (raw Scopus data at the micro-level of individual scientists). In a cohort-based and longitudinal fashion, we follow individual men and women scientists over time, from their first to their last publication (N=86,178). We examine four academic disciplines: AGRI (agricultural and biological sciences), BIO (biochemistry, genetics, and molecular biology), IMMU (immunology and microbiology), and NEURO (neuroscience). We apply survival analysis, conceptualizing scientific life as a sequence of scholarly publishing events. Our Kaplan-Meier survival analysis shows how women disappear from science: in BIO, about 60% are still in science after 5 years, 40% after 10 years, and only 20% by the end of the period examined (i.e., after 19 years). The percentages are substantially higher for men: approximately 70%, 50%, and 30%, respectively. Kaplan-Meier estimations indicate that women in the largest discipline (BIO) are 23.26% more likely to leave science after 10 years and 39.74% more likely to leave science at the end of the study period. Gender difference in attrition, slightly visible after 5 years, increase consistently in later career stages. The probability of surviving for women after 15 years varies considerably, from 47.8% in AGRI to 27.6% in IMMU; for men, the probability is about a fifth higher. Our data show that with the passage of time, women disappear from science in ever-larger proportions compared to men. Gender differences in attrition in the four disciplines have been and continue to be high, but comparing the 2000 and 2010 cohorts, have slightly decreased over time.

scientific communication and education↗

bioRxiv2025

Martini 3 protein models - a practical introduction to different structure bias models and their comparison

Biophysical characterization of protein structure and dynamics is essential in many scientific fields, including molecular biology, drug discovery, and enzyme design. Molecular dynamics (MD) simulations have become an increasingly important tool for studying these properties. This chapter provides a hands-on introduction to protein modeling with the Martini 3 coarse-grained (CG) force field. We outline its two-layer framework, where the first layer defines molecular topology and interactions, while the second layer applies structural bias to maintain secondary, tertiary, and quaternary structures. Three structure bias approaches - Elastic Network (EN), G[o]Martini, and OLIVES - are discussed, highlighting their advantages and trade-offs. Using a protein kinase as a case study, we demonstrate the step-by-step setup of Martini 3 protein models for simulations with the program package GROMACS, including system preparation, fine-tuning, and validation against atomistic reference simulations. Additionally, we explore the combination of an intrinsically disordered region (IDR) with a structured protein domain. By the end of this chapter, readers will have the necessary expertise to apply Martini 3-based protein modeling to a wide range of research applications.

biophysics↗

bioRxiv2017

Wx: a neural network-based feature selection algorithm for next-generation sequencing data

MotivationNext-generation sequencing (NGS), which allows the simultaneous sequencing of billions of DNA fragments simultaneously, has revolutionized how we study genomics and molecular biology by generating genome-wide molecular maps of molecules of interest. For example, an NGS-based transcriptomic assay called RNA-seq can be used to estimate the abundance of approximately 190,000 transcripts together. As the cost of next-generation sequencing sharply declines, researchers in many fields have been conducting research using NGS. The amount of information produced by NGS has made it difficult for researchers to choose the optimal set of target genes (or genomic loci).\n\nResultsWe have sought to resolve this issue by developing a neural network-based feature (gene) selection algorithm called Wx. The Wx algorithm ranks genes based on the discriminative index (DI) score that represents the classification power for distinguishing given groups. With a gene list ranked by DI score, researchers can institutively select the optimal set of genes from the highest-ranking ones. We applied the Wx algorithm to a TCGA pan-cancer gene-expression cohort to identify an optimal set of gene-expression biomarker (universal gene-expression biomarkers) candidates that can distinguish cancer samples from normal samples for 12 different types of cancer. The 14 gene-expression biomarker candidates identified by Wx were comparable to or outperformed previously reported universal gene expression biomarkers, highlighting the usefulness of the Wx algorithm for next-generation sequencing data. Thus, we anticipate that the Wx algorithm can complement current state-of-the-art analytical applications for the identification of biomarker candidates as an alternative method.\n\nAvailabilityhttps://github.com/deargen/DearWX\n\nContactkangk1204@dankook.ac.kr\n\nSupplementary informationSupplementary data are available at online.

bioinformatics↗

bioRxiv2026

High-resolution dissection of concept acquisition in different families of protein language models

Protein language models have been increasingly successful on tasks ranging from fitness prediction to functional design, yet what biological knowledge they acquire and where it is encoded within their internal representations remain underexplored. Through a high-resolution layer-by-layer interpretability analysis of 8 models from the ESM2 and AMPLIFY families on 22 concepts from human proteome annotations, we found that these models encode concepts of increasing levels of complexity along their depth: basic physicochemical properties and linear motifs are best captured by early-layer embeddings, secondary structure from subsequent layers, and domain-level semantics from middle layers. Principal component projections of these embeddings showed that they separate biologically meaningful protein groupings, and molecular-biology-inspired interventions demonstrated that pLM embeddings can discriminate phosphomimic-active from inactive mutants. Perhaps surprisingly, we observed that pretraining data and compute had a greater impact on the linear emergence of biological concepts than scaling up parameters. By revealing where biological knowledge is captured in pLMs and which choices shape its emergence, our work offers insights to develop more robust, biologically grounded protein language models.

bioinformatics↗

bioRxiv2020

Probabilistic Framework for Integration of Mass Spectrum and Retention Time Information in Small Molecule Identification

MotivationIdentification of small molecules in a biological sample remains a major bottleneck in molecular biology, despite a decade of rapid development of computational approaches for predicting molecular structures using mass spectrometry (MS) data. Recently, there has been increasing interest in utilizing other information sources, such as liquid chromatography (LC) retention time (RT), to improve the MS based identifications. ResultsWe put forward a probabilistic modelling framework to integrate MS and RT data of multiple features in an LC-MS experiment. We model the MS measurements and all pairwise retention order information as a Markov random field and use efficient approximate inference for scoring and ranking potential molecular structures. Our experiments show improved identification accuracy by combining tandem mass spectrometry data (MS2) and retention orders using our approach, thereby outperforming state-of-the-art methods. Furthermore, we demonstrate the benefit of our model when only a subset of LC-MS features have MS2 measurements available besides MS1. Availability and implementationSoftware and data is freely available at https://github.com/aalto-ics-kepaco/msms_rt_score_integration. Contacteric.bach@aalto.fi

systems biology↗

bioRxiv2022

Electricity-free nucleic acid extraction method from dried blood spots on filter paper for point-of-care diagnostics

BackgroundNucleic acid extraction is a crucial step for molecular biology applications, being a determinant for any diagnostic test procedure. Dried blood spots (DBS) have been used for decades for serology, drug monitoring, environmental investigations, and molecular studies. Nevertheless, nucleic acid extraction from DBS remains one of the main challenges to translate them to the point-of-care (POC). MethodWe have developed a fast nucleic acid extraction (NAE) method from DBS which is electricity-free and relies on cellulose filter papers (DBSFP). The performance of NAE was assessed with loop-mediated isothermal amplification (LAMP), targeting the human reference gene beta-actin. The developed method was evaluated against FTA cards and magnetic bead-based purification, using time-to-positive (min) for comparative analysis. We optimised and validated the developed method for elution (eluted disk) and disk directly in the reaction (in-situ disk), RNA and DNA detection, and whole blood stored in anticoagulants (K2EDTA and lithium heparin). Furthermore, the compatibility of DBSFP with colourimetric detection was studied to show the transferability to the POC. ResultsThe proposed DBSFP is based on grade 3 filter paper pre-treated with 8% (v/v) igepal surfactant, 1 min washing step with PBS 1X and elution in TE 1X buffer after 5 min incubation at room temperature, enabling NAE under 7 min. Obtained results were comparable to gold standard methods across tested matrices, targets and experimental conditions, demonstrating the versatility of the methodology. Lastly, eluted disk colourimetric detection was achieved with a sample-to-result turnaround time under 35 min. ConclusionsThe developed method is a fast, electricity-free, and low-cost solution for NAE from DBSFP enabling molecular testing in virtually any POC setting.

molecular biology↗

bioRxiv2022

Mathematical Modeling Of Systems Biology

A modeling is a mathematical tool, like a microscope, which allows consequences to logically follow from a set of assumptions by which a real world problem can be described by a mathematical formulation. It has become indispensable tools for integrating and interpreting heterogeneous biological data, validating hypothesis and identifying potential diagnostic markers. The modern molecular biology that is characterized by experiments that reveal the behaviours of entire molecular systems is called systems biology. A fundamental step in synthetic biology and systems biology is to derive appropriate mathematical model for the purposes of analysis and design. This manuscript has been engaged in the use of mathematical modeling in the Gene Regulatory System (GRN). Different mathematical models that are inspired in gene regulatory network such as Central dogma, Hill function, Gillespie algorithm, Oscillating gene network and Deterministic vs Stochastic modelings are discussed along with their codes that are programmed in Python using different modules. Here, we underlined that the model should describes the continuous nature of the biochemical processes and reflect the non-linearity. It is also found that the stochastic model is far better than deterministic model to calculate future event exactly with low chance of error.

systems biology↗

bioRxiv2022

Nanopore sequencing unveils the complexity of the cold-activated murine brown adipose tissue transcriptome

1.Alternative transcription increases transcriptome complexity by expression of multiple transcripts per gene and thus fine tunes cellular identity and function. Annotation and quantification of transcripts at complex loci using short-read sequencing is non-trivial. Recent long-read sequencing methods such as those from Oxford Nanopore Technologies (ONT) and Pacific Biosciences aim at overcoming these problems by sequencing full length transcripts. Activation of BAT thermogenesis involves major transcriptomic remodelling and positively affects metabolism via increased energy expenditure and endocrine factors. Here we comprehensively benchmark features of ONT long-read sequencing protocols compared to Illumina shortread sequencing assessing alignment characteristics, gene and transcript detection and quantification, differential gene and transcript expression, transcriptome reannotation and differential transcript usage (DTU). We find that ONT sequencing is superior to Illumina for transcriptome reassembly and reduces the risk of false-positive events due to the ability to unambiguously map reads to transcripts, at the expense of statistical power for calling differentially expressed features. We identified novel isoforms of genes undergoing DTU in cold-activated BAT including Cars2, Adtrp, Acsl5, Scp2, Aldoa and Pde4d, validated by RT-qPCR. Finally, we provide a reannotation of the murine iBAT transcriptome as a valuable resource for researchers interested in the molecular biology underlying the regulation of BAT.

molecular biology↗

bioRxiv2025

Quantitative Assessment of Protein-DNA Interactions via SYBR Green Fluorescence

Protein-DNA interactions are crucial for cellular processes, but current quantification methods lack sensitivity. We developed the Real-time PCR-based DNA Binding Assay (RP-DBA) to detect and quantify these interactions. The target protein, expressed as a Strep-tag II fusion, is purified and incubated with double-stranded DNA probes containing 4 bp 3 overhangs. Protein-DNA complexes are immobilized on Strep-Tactin beads, washed, and eluted. A complementary single-stranded DNA amplification arm is added, extended by Taq polymerase, and quantified via qPCR with SYBR Green. RP-DBA enables real-time kinetic analysis and is 4- to 10-fold more sensitive than EMSA, depending on amplification arm length (30-90 bp). Its simplicity, speed, accuracy, and high-throughput potential make it a valuable tool for advancing molecular biology research.

molecular biology↗

bioRxiv2024

The Leader of the Capsid protein from Feline calicivirus must be palmitoylated and form oligomers through disulfide bonds for efficient viral replication

Feline calicivirus (FCV), a member of the Vesivirus genus and a model for studying of the members of the Caliciviridae, has been used to understand the biology and viral replication of etiological agents of medical and veterinary importance. The leader of the capsid (LC) protein is exclusive to the Vesivirus genus members whose importance for a successful viral replication has been demonstrated; however, its cell localization, post-translational modifications, and specific functions throughout the viral replication cycle are poorly understood. We have determined that the expression of the LC protein from FCV in a virus-free system is located at the mitochondria and induces apoptosis; furthermore, the in vitro characterization of a purified LC protein showed that it has viroporin characteristics. Here, using an LC-reactive serum, the LC protein expression kinetics, cellular localization, post-translational modifications, and association with cellular PDI proteins in FCV infected cells was determined. We found that the LC protein is present on the membrane of infected cells and in the supernatants, independently of cell lysis, which suggests that it is actively secreted. Moreover, we found that LC protein is palmitoylated during infection and that palmitoylation inhibition alters its levels and subcellular localization. Finally, we determined that the LC protein forms oligomers dependent on disulfide bonds mediated by PDI activity. In addition, it interacts with PDIA3, a disulfide isomerase known as an important factor in the replication of several viruses. The results indicate that the LC protein from FCV might have multiple roles during FCV replication. IMPORTANCEFeline calicivirus (FCV) is a highly transmissible virus that represents a significant cause of upper respiratory infection in domestic and wild cats worldwide. FCV also serves as one of the most valuable models for studying calicivirus biology, as unlike most members of the family, known to cause diseases in animals and humans, it can be easily grown in cell culture. Since there are no efficacious vaccines or antivirals against most caliciviruses, studying their molecular biology and the relationship between viral and cellular components is essential for developing strategies for their prevention and control.

molecular biology↗

bioRxiv2025

Mechanistic Investigation of LncRNA-ZMIZ1-AS1 in the Malignant Phenotype of Hepatocellular Carcinoma

BackgroundHepatocellular carcinoma (HCC) is one of the most fatal cancers worldwide, with high recurrence and mortality rates. LncRNA-ZMIZ1-AS1 has attracted considerable attention for its role in the tumor microenvironment, particularly in the regulation of HCC cell growth, invasion, and biological behavior. However, the specific mechanisms of LncRNA-ZMIZ1-AS1 in HCC remain unclear. MethodsWe analyzed the relationship between LncRNA-ZMIZ1-AS1 and HCC clinical prognosis using bioinformatics with GEO and TCGA datasets. We also conducted molecular biology experiments, including cell culture, real-time PCR, and Western blotting, as well as cell viability and flow cytometry analyses, to investigate the effects of LncRNA-ZMIZ1-AS1 knockdown on HCC cell proliferation, migration, invasion, and apoptosis. Furthermore, we used Gene Set Enrichment Analysis (GSEA) to identify potential signaling pathways involving LncRNA-ZMIZ1-AS1. ResultsLncRNA-ZMIZ1-AS1 was highly expressed in HCC patients and significantly associated with clinical characteristics, such as gender and clinical stage. Knockdown of LncRNA-ZMIZ1-AS1 notably inhibited HCC cell proliferation, migration, and invasion while increasing apoptosis rates. Additionally, GSEA suggested a potential association between high LncRNA-ZMIZ1-AS1 expression and the NOTCH signaling pathway. ConclusionThe high expression of LncRNA-ZMIZ1-AS1 in HCC likely promotes malignant phenotypes by regulating signaling pathways such as NOTCH. This finding underscores its potential as a biomarker for HCC, providing a theoretical basis for further research on its molecular mechanisms and clinical applications in liver cancer.

molecular biology↗

bioRxiv2026

Purification of filamentous phage ssDNA genomes using Chaotropic salt/silica methods

Filamentous phages play important roles in molecular biology and bacterial pathogenesis. There is therefore great interest in the genomes of the phages, including M13 and Pf, which infect Escherichia coli and Pseudomonas aeruginosa, respectively. However, efficient methods for purifying single-stranded DNA (ssDNA) from filamentous phages are scarce and rely mainly on repurposed commercial kits developed for dsDNA. These kits yield variable amounts of ssDNA, varying considerably from one phage to another, their solutions are proprietary and troubleshooting, and customizing the protocol to improve purification is not possible. Here, we developed and optimized two protocols based on ssDNA binding to silica columns in the presence of guanidine hydrochloride and potassium iodide chaotropic salts, for filamentous phages M13KO7 and Pf4. The protocols achieved 60 to 100% recovery, equal to or higher than the Monarch Spin gDNA Extraction and DNeasy Blood and Tissue kits, with similar A260/280 and A260/230 ratios, intact DNA in agarose gels, and no detectable host genomic DNA by PCR. Recovery depended critically on the chaotropic salts and on ethanol, and did not increase at lower pH, consistent with a water-exclusion model in which hydrophobic interactions and hydrogen bonding dominate over electrostatic repulsion. Phage-specific differences in recovery indicate that optimization may be needed for some phages, which the fully customizable nature of these protocols facilitates. Comparing the virion ssDNA with the double-stranded replicative form (RF) revealed genome variants in both phages, including a shorter M13KO7 form lacking the kanamycin resistance-p15A insert, and illustrates the value of analyzing both RF and ssDNA forms to study filamentous phage biology. Implementing these protocols in routine filamentous phage workflows should facilitate their characterization and engineering.

molecular biology↗