title: Identification of unique essential proteins from a Mycobacterium tuberculosis F15/LAM4/KZN phage secretome library.
abstract: Mycobacterium tuberculosis (Mtb) is the causative agent of tuberculosis disease (TB), the leading cause of death from bacterial infection worldwide. Although treatable, the resurgence of multidrug-resistant and extensively drug-resistant TB is a major setback for the fight against TB globally. Consequently, there is an urgent need for new Mtb-derived biomarkers for use in the design of new drugs and rapid point-of-care diagnostic or prognostic tools for the management of TB transmission. Therefore, the present study aimed to identify unique Mtb-secreted proteins from the extensively drug-resistant Mtb F15/LAM4/KZN phage secretome library. A whole genome library was constructed using genomic DNA fragments of the Mtb F15/LAM4/KZN strain. A phage secretome sub-library of 8 × 103 clones was prepared and phage DNA was sequenced from 120 randomly selected clones. DNA sequence BLAST analysis identified 86 open reading frames. Using bioinformatics tools and databases, 10 proteins essential for in vivo growth and survival of Mtb (Nrp, PssA, MmpL5, SirA, GatB, EspA, TopA, EccCa1, Rv1634 and Rv3103c) were identified. Proteins essential for the growth and survival of Mtb during infection have potential application in the development of diagnostic tools, new drugs and vaccines. Further studies will be conducted to evaluate their potential application in the fight against TB.
authors.affiliation: Discipline of Microbiology, School of Life Sciences, College of Agriculture, Engineering and Science, University of KwaZulu-Natal, Westville 3630, South Africa.
authors.forename: Thamsanqa Emmanuel
authors.initials: TE
authors.lastname: Chiliza
authors.affiliation: Medical Microbiology and Infection Control, School of Laboratory Medicine and Medical Sciences, College of Health Sciences, University of KwaZulu-Natal, Durban, South Africa.
authors.forename: Manormoney
authors.initials: M
authors.lastname: Pillay
authors.affiliation: Discipline of Microbiology, School of Life Sciences, College of Agriculture, Engineering and Science, University of KwaZulu-Natal, Westville 3630, South Africa.
authors.forename: Balakrishna
authors.initials: B
authors.lastname: Pillay
mesh_terms: D001426:Bacterial Proteins / Q000378:metabolism*; D019295:Computational Biology / Q000379:methods; D005814:Genes, Viral; D016680:Genome, Bacterial; D015698:Genomic Library; D058977:Molecular Sequence Annotation; D009160:Mycobacteriophages / Q000502:physiology*; D009169:Mycobacterium tuberculosis / Q000235:genetics / Q000378:metabolism* / Q000821:virology*; D021382:Protein Sorting Signals; D021381:Protein Transport; D020543:Proteome; D040901:Proteomics / Q000379:methods; D014764:Viral Proteins / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins; D021382:Protein Sorting Signals; D020543:Proteome; D014764:Viral Proteins
keywords: M. tuberculosis; biomarkers; diagnosis; phage display; secreted proteins; vaccine