Preparation a Recombinant Form of Pneumolysin Protein from Streptococcus pneumoniae.
Streptococcus pneumoniae
affinity chromatography
nucleotide sequence
recombinant pneumolysin
sequencing
Journal
Bulletin of experimental biology and medicine
ISSN: 1573-8221
Titre abrégé: Bull Exp Biol Med
Pays: United States
ID NLM: 0372557
Informations de publication
Date de publication:
Apr 2023
Apr 2023
Historique:
received:
09
10
2022
medline:
22
5
2023
pubmed:
10
5
2023
entrez:
10
5
2023
Statut:
ppublish
Résumé
A recombinant form of pneumolysin from Streptococcus pneumoniae was obtained. By using Vector NTI Advance 11.0 bioinformatic analysis software, specific primers were designed in order to amplify the genome fragment of strain No. 3358 S. pneumoniae serotype 19F containing the nucleotide sequence encoding the full-length pneumolysin protein. A PCR product with a molecular weight corresponding to the nucleotide sequence of the S. pneumoniae genome fragment encoding the full-length pneumolysin was obtained. An expression system for recombinant pneumolysin in E. coli was constructed. Sequencing confirmed the identity of the inserted nucleotide sequence encoding the full-length recombinant pneumolysin synthesized in E. coli M15 strain. Purification of the recombinant protein was performed by affinity chromatography using Ni-Sepharose in 8 M urea buffer solution. Confirmation of the recombinant protein was performed by immunoblotting with monoclonal antibodies to pneumolysin.
Identifiants
pubmed: 37160796
doi: 10.1007/s10517-023-05785-3
pii: 10.1007/s10517-023-05785-3
doi:
Substances chimiques
plY protein, Streptococcus pneumoniae
0
Streptolysins
0
Bacterial Proteins
0
Recombinant Proteins
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
749-753Informations de copyright
© 2023. Springer Science+Business Media, LLC, part of Springer Nature.
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