About: A simpler and more cost-effective peptide biosynthetic method using the truncated GST as carrier for epitope mapping

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About: A simpler and more cost-effective peptide biosynthetic method using the truncated GST as carrier for epitope mapping Goto Sponge NotDistinct Permalink

An Entity of Type : schema:ScholarlyArticle, within Data Space : wasabi.inria.fr associated with source document(s)

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type	Academic Article research paper schema:ScholarlyArticle
isDefinedBy	Covid-on-the-Web dataset
title	A simpler and more cost-effective peptide biosynthetic method using the truncated GST as carrier for epitope mapping
Creator	Wang, Jian Chen, Ling-Han Gu, Shao-Hua Ji, Chao-Neng Lian, Wen-Bo Tang, Hai-Ping Xie, Yi Xu, Wan-Xiang Zhan, Jian-Min Gupta, Satish
source	Medline; PMC
abstract	There is a need to develop better methods for epitope mapping and/or identification of antibody-recognizing motifs. Here, we describe improved biosynthetic peptide (BSP) method using a newly developed plasmid pXXGST-3 as vector, which has a viral E7 gene in the cloning sites of pXXGST-1. It is crucial to employ pXXGST-3 instead of pXXGST-1, since it makes use of the BSP method simpler and easier to perform, and more cost-effective for epitope mapping. These merits are embodied in two aspects: i) convenient recovery of double enzyme-digested product due to the existence of 315 bp inserted between BamH I and Sal I sites, and thus greatly reducing the production of self-ligation clones, and ii) no longer requiring control protein when screening recombinant (r-) clones expressing 8/18mer peptides by running polyacrylamide gel electrophoresis. The protocol involves the following core steps: (i) design of plus and minus strands of DNA fragments encoding overlapping 8/18mer peptides; (ii) chemical synthesis of the designed DNA fragments; (iii) development of r-clones using pXXGST-3 vector expressing each 8/18mer peptide fused with truncated GST188 protein; (iv) screening r-clones by running the cell pellets from each induced clone on SDS-PAGE gel followed by sequencing of inserted DNA fragments for each verified r-clone; and (v) Western blotting with either monoclonal antibodies or polyclonal antibodies. This improved GST188-BSP method provides a powerful alternative tool for epitope mapping.
has issue date	2017-10-12(xsd:dateTime)
bibo:doi	10.1371/journal.pone.0186097
bibo:pmid	29023483
has license	cc-by
sha1sum (hex)	e408e143da8a102c318c2d30be909628b96e1175
schema:url	https://doi.org/10.1371/journal.pone.0186097
resource representing a document's title	A simpler and more cost-effective peptide biosynthetic method using the truncated GST as carrier for epitope mapping
has PubMed Central identifier	PMC5638316
has PubMed identifier	29023483
schema:publication	PLoS One
resource representing a document's body	covid:e408e143da8a102c318c2d30be909628b96e1175#body_text
is schema:about of	named entity 'peptide' named entity 'Lian' named entity 'screening' named entity 'reducing' covid:arg/e408e143da8a102c318c2d30be909628b96e1175 named entity 'BSP' named entity 'peptides' named entity 'protein' named entity 'epitope mapping' named entity 'cloning' named entity 'core' named entity 'inserted' named entity 'motifs' named entity 'ACCESS' named entity 'gene' named entity 'This' named entity 'peptide' named entity 'protein' named entity 'vector' named entity 'enzyme' named entity 'chemical synthesis' named entity 'epitope mapping' named entity 'doi' named entity 'Western blotting' named entity 'SDS-PAGE' named entity 'protein' named entity 'OPEN ACCESS' named entity 'clones' named entity 'clones' named entity 'peptide' named entity 'enzyme' named entity 'gene' named entity 'Stv' named entity 'annealed' named entity 'SDS-PAGE' named entity 'laboratory animal' named entity 'DNA fragments' named entity 'protein' named entity 'mAb' named entity 'antigenic' named entity 'Centrifuge' named entity 'target gene' named entity 'Western blotting' named entity 'proteolytic' named entity 'clones' named entity 'clone' named entity 'Shanghai' named entity 'antibodies' named entity 'Western blotting' named entity 'phosphate-buffered saline' named entity 'nitrocellulose' named entity 'cross-reactive' named entity 'clones' named entity 'peptides' named entity 'peptide' named entity 'fusion proteins' named entity 'Chemical synthesis' named entity 'sequence alignment' named entity 'protein' named entity 'G-3' named entity 'homologous proteins' named entity 'DNA fragments' named entity 'antigenic peptides' named entity 'cell lysates' named entity 'clones' named entity 'mAb' named entity 'oligonucleotide' named entity 'C-terminus' named entity 'agarose gel electrophoresis' named entity 'motif' named entity 'peptides' named entity 'clones'

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