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1860797892721115136
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INTELEK Repository
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Online Access
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https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection407072
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2024-08-26 16:18:35
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Terengganu
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Restricted Document
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14559
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UniSZA
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2519-01-FH03-FBIM-19-30933.pdf
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| person |
Fatimah Hashim
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oai_dc
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https://intelek.unisza.edu.my/intelek/pages/view.php?ref=14559
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14559 https://intelek.unisza.edu.my/intelek/pages/view.php?ref=14559 https://intelek.unisza.edu.my/intelek/pages/search.php?search=!collection407072 Restricted Document Conference Conference Paper application/pdf Adobe Acrobat Pro DC 20 Paper Capture Plug-in with ClearScan 1.7 Fatimah Hashim 2024-08-26 16:18:35 125 2519-01-FH03-FBIM-19-30933.pdf UniSZA Private Access Development of CBM-Fusion Protein with Oligomerization Domain (OD) from Pseudomonas aeruginosa Pseudaminidase Carbohydrate binding module (CBM) from Vibrio cholerae sialidase is located at the N-terminal region which flank the central catalytic domain of the protein. It is belong to the Family 40 which recognizes single sialic acid moiety as the binding ligand. Protein usually tend to have weaker binding affinity towards its substrate, therefore a multivalent approach was carried out in order to enhance protein binding affinity via an avidity effect. In this study, DNA fragment encoding the CBM40 from V. cholerae sialidase was amplified by PCR method before cloning in E. coli BL21(DE3) using pET-22(b+) as a vector. The gene composed of 174 amino acids from an open reading frame comprising 522 bp nucleotide sequence. CBM40-fusion protein was developed by linking the second domain known as Oligomerization domain (OD) which isolate from Pseudomonas aeruginosa pseudaminidase. The domain consists of an open reading frame of 321 bp with the ability to form a trimeric protein molecule. The OD domain was amplified by PCR method with the addition of BamH1 and EcoR1 sites before ligation was performed on pET-22b(+) vector. In addition, five amino acids linker length was generated to flank in the CBM and the OD domain. Choices of fusion linkers were designed composed of flexible residues like glycine (G) and serine (S) which are flexible group of amino acids. This recombinant fusion construct was cloned into the pET-22b(+) vector and transform into E. coli BL21 (DE3) host. Positive recombinant fusion molecule was identified by colony PCR and confirmed by DNA sequencing techniques. A comprehensive characterization of the recombinant fusion proteins is currently under evaluation. This CBM-OD construct has a big potential to be developed as a therapeutics agent to combat harmful pathogens which recognize sialic acid as the binding substrate. 77 2 nd National Seminar on Security and Sustainability Biology (BIOSES) 2019 Terengganu
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| spellingShingle |
Development of CBM-Fusion Protein with Oligomerization Domain (OD) from Pseudomonas aeruginosa Pseudaminidase
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| summary |
Carbohydrate binding module (CBM) from Vibrio cholerae sialidase is located at the N-terminal region which flank the central catalytic domain of the protein. It is belong to the Family 40 which recognizes single sialic acid moiety as the binding ligand. Protein usually tend to have weaker binding affinity towards its substrate, therefore a multivalent approach was carried out in order to enhance protein binding affinity via an avidity effect. In this study, DNA fragment encoding the CBM40 from V. cholerae sialidase was amplified by PCR method before cloning in E. coli BL21(DE3) using pET-22(b+) as a vector. The gene composed of 174 amino acids from an open reading frame comprising 522 bp nucleotide sequence. CBM40-fusion protein was developed by linking the second domain known as Oligomerization domain (OD) which isolate from Pseudomonas aeruginosa pseudaminidase. The domain consists of an open reading frame of 321 bp with the ability to form a trimeric protein molecule. The OD domain was amplified by PCR method with the addition of BamH1 and EcoR1 sites before ligation was performed on pET-22b(+) vector. In addition, five amino acids linker length was generated to flank in the CBM and the OD domain. Choices of fusion linkers were designed composed of flexible residues like glycine (G) and serine (S) which are flexible group of amino acids. This recombinant fusion construct was cloned into the pET-22b(+) vector and transform into E. coli BL21 (DE3) host. Positive recombinant fusion molecule was identified by colony PCR and confirmed by DNA sequencing techniques. A comprehensive characterization of the recombinant fusion proteins is currently under evaluation. This CBM-OD construct has a big potential to be developed as a therapeutics agent to combat harmful pathogens which recognize sialic acid as the binding substrate.
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| title |
Development of CBM-Fusion Protein with Oligomerization Domain (OD) from Pseudomonas aeruginosa Pseudaminidase
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| title_full |
Development of CBM-Fusion Protein with Oligomerization Domain (OD) from Pseudomonas aeruginosa Pseudaminidase
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| title_fullStr |
Development of CBM-Fusion Protein with Oligomerization Domain (OD) from Pseudomonas aeruginosa Pseudaminidase
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| title_full_unstemmed |
Development of CBM-Fusion Protein with Oligomerization Domain (OD) from Pseudomonas aeruginosa Pseudaminidase
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| title_short |
Development of CBM-Fusion Protein with Oligomerization Domain (OD) from Pseudomonas aeruginosa Pseudaminidase
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| title_sort |
development of cbm-fusion protein with oligomerization domain (od) from pseudomonas aeruginosa pseudaminidase
|