Identification and molecular characterization of plasmid-mediated verotoxin gene in non-o157 escherichia coli / (Record no. 96288)

MARC details
000 -LEADER
fixed length control field 04890nam a2200349 i 4500
003 - CONTROL NUMBER IDENTIFIER
control field MY-KuUP
005 - DATE AND TIME OF LATEST TRANSACTION
control field 20251125105944.0
006 - FIXED-LENGTH DATA ELEMENTS--ADDITIONAL MATERIAL CHARACTERISTICS
fixed length control field a||||fr|||| 001 0
007 - PHYSICAL DESCRIPTION FIXED FIELD--GENERAL INFORMATION
fixed length control field ta
008 - FIXED-LENGTH DATA ELEMENTS--GENERAL INFORMATION
fixed length control field 220304t20212021my a|||fram|| 001 0 eng d
020 ## - INTERNATIONAL STANDARD BOOK NUMBER
International Standard Book Number THE0009239(Local)
040 ## - CATALOGING SOURCE
Original cataloging agency UMP
Language of cataloging eng
Transcribing agency UMP
Description conventions rda
090 ## - LOCALLY ASSIGNED LC-TYPE CALL NUMBER (OCLC); LOCAL CALL NUMBER (RLIN)
Classification number (OCLC) (R) ; Classification number, CALL (RLIN) (NR) FIST .F39 2021 r Thesis
100 1# - MAIN ENTRY--PERSONAL NAME
Personal name Md. Fazlul Karim Khan,
Relator term author.
245 10 - TITLE STATEMENT
Title Identification and molecular characterization of plasmid-mediated verotoxin gene in non-o157 escherichia coli /
Statement of responsibility, etc. Md. Fazlul Karim Khan
264 #1 - PRODUCTION, PUBLICATION, DISTRIBUTION, MANUFACTURE, AND COPYRIGHT NOTICE
Place of production, publication, distribution, manufacture Kuantan, Pahang :
Name of producer, publisher, distributor, manufacturer UMP,
Date of production, publication, distribution, manufacture, or copyright notice 2021
264 #4 - PRODUCTION, PUBLICATION, DISTRIBUTION, MANUFACTURE, AND COPYRIGHT NOTICE
Place of production, publication, distribution, manufacture © 2021
300 ## - PHYSICAL DESCRIPTION
Extent xxi, 241 pages :
Other physical details illustrations (some color) ;
Dimensions 30 cm. +
Accompanying material 1 CD ROM
336 ## - CONTENT TYPE
Content type term text
Source rdacontent
337 ## - MEDIA TYPE
Media type term unmediated
Source rdamedia
338 ## - CARRIER TYPE
Carrier type term volume
Source rdacarrier
347 ## - DIGITAL FILE CHARACTERISTICS
File type text file
Encoding format PDF
Source rda
500 ## - GENERAL NOTE
General note Faculty of Industrial Sciences and Technology
502 ## - DISSERTATION NOTE
Dissertation note Thesis (Doctor of Philosophy) -- Universiti Malaysia Pahang – 2021
504 ## - BIBLIOGRAPHY, ETC. NOTE
Bibliography, etc. note Includes bibliographical references
520 3# - SUMMARY, ETC.
Summary, etc. Diarrheagenic non-O157 verotoxin-producing Escherichia coli (VTEC) associates with endemic infantile diarrhoea, causing morbidity and mortality across the world. Thus, the existence and prevalence of non-O157 VTEC have emerged as a particular public health concern. This study investigated the biological activities of the plasmid-mediated Vtx gene in non-O157 VTEC isolates. Besides, the extracted Vtx gene sequence information was used in tertiary structure modelling followed by thermostability analysis as a computational approach of potential vaccine candidates. Standard microbiological techniques identified non-repeated, clinically significant E. coli isolates. Serological assays discerned non-O157 E. coli serogroups while all these isolates were subjected to a polymerase chain reaction for investigating virulent genes, Vtx1 and Vtx2. Subsequently, the Close and Rodriguez method and Perfectprep plasmid mini preparation kit were used to detect plasmid harbored isolates. These plasmid harbored isolates were further investigated using disk diffusion assay, phage induction, and plasmid curing followed by polymerase chain reaction. Meanwhile, the sequence of the Vtx gene was analyzed using several bioinformatics tools, including BioEdit, NCBI-BLAST, ClustalW-X, and MEGA 5.0, etc. Furthermore, most potential Vtx gene sequences were modelled using MODELLER and refined via molecular dynamics simulations using GROMACS. Finally, the Vtx gene as a potential vaccine candidate was evaluated by incorporating scales of beta turns, hydropathicity, surface accessibility, and antigenicity in MATLAB. In this cross-sectional study, 137 clinical isolates of E. coli were collected from Hospital Tengku Ampuan Afzan, Kuantan, Malaysia. Different PCR sets characterized 49 clinical isolates of sorbitol positive non-O157 E. coli while twenty-nine isolates harbored Vtx genes (Vtx1 and Vtx2). Among 29 (59.18 %) strains of Vtx producing non-O157 E. coli¸ Vtx1 in 21 isolates (42.85 %), Vtx2 in 5 isolates (10.20 %) and both Vtx1 and Vtx2 was observed in 3 (6.12 %) isolates. Besides that, 26 multidrugresistant isolates possessed single-sized and multiple plasmids with different molecular weight varied from 1.8 kb to 7.6 kb. Furthermore, eight isolates changed their susceptibility patterns, while three isolates were found to lose plasmid after post plasmid curing treatment, and the rest of the 15 isolates remained unchanged. Finally, different PCR sets characterized 3 plasmid-mediated Vtx gene producing non-O157 E. coli while the absence of lysogenic phages indicated that Vtx genes were not phage mediated. On the other hand, these three plasmids mediated Vtx gene in non-O157 VTEC isolates were used as a template DNA for sequences of interest. The sequencing results confirmed the occurrence of Vtx genes and demonstrated higher similarity in nucleotides level. Moreover, Verify 3D, ERRAT, and PROCHECK showed that the generated Vtx1-1 protein model has a high stereochemical quality. In addition, structural analysis of the thermal profile demonstrated the stability of Vtx1-1 models structure at 40 °C. As the temperature rises, the stability of the Vtx1-1 protein model was disrupted due to flexible secondary loops as well as the presence of non-charged and hydrophilic amino acid residues. However, DSGTGDNLF (47-55) and MIDSGTGDN (45-53), two 9-mer peptides with a score of 1.242 and 1.229 respectively, could be the most effective B-cell epitopes and a possible vaccine candidate against VTEC infections. The information obtained from this study is expected to improve the molecular synthesis applications in the future.
610 20 - SUBJECT ADDED ENTRY--CORPORATE NAME
Corporate name or jurisdiction name as entry element Faculty of Industrial Sciences and Technology
General subdivision Dissertations
650 #0 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element Universities and colleges
General subdivision Dissertations
650 #0 - SUBJECT ADDED ENTRY--TOPICAL TERM
Topical term or geographic name entry element Theses
942 ## - ADDED ENTRY ELEMENTS (KOHA)
Source of classification or shelving scheme Library of Congress Classification
Koha item type Restricted Collection
Holdings
Withdrawn status Lost status Source of classification or shelving scheme Damaged status Use restrictions Not for loan Collection Home library Current library Shelving location Date acquired Total checkouts Full call number Barcode Date last seen Copy number Price effective from Koha item type
  Not lost Library of Congress Classification   Restricted access Not for loan Reference UMPLIB GAMBANG UMPLIB GAMBANG Reference 12/01/2022   FIST .F39 2021 r Thesis T000001650 12/05/2022 1 12/01/2022 Restricted Collection
  Not lost Library of Congress Classification   Restricted access Not for loan Reference UMPLIB GAMBANG UMPLIB GAMBANG   04/03/2022   CD 12991 T000001651 30/01/2023 1 04/03/2022 Restricted Collection

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