Progress toward Plug-and-Play Polymer Strings for Optical Tweezers Experiments: Concatenation of DNA Using Streptavidin Linkers

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dc.citation.issue7
dc.citation.volume7
dc.contributor.authorMohandas N
dc.contributor.authorKent LM
dc.contributor.authorRaudsepp A
dc.contributor.authorJameson GB
dc.contributor.authorWilliams MAK
dc.coverage.spatialUnited States
dc.date.accessioned2023-11-01T23:39:15Z
dc.date.accessioned2023-11-03T04:45:40Z
dc.date.available2022-02-10
dc.date.available2023-11-01T23:39:15Z
dc.date.available2023-11-03T04:45:40Z
dc.date.issued2022-02-22
dc.description.abstractStreptavidin is a tetrameric protein that is renowned for its strong binding to biotin. The robustness and strength of this noncovalent coupling has led to multitudinous applications of the pairing. Within the streptavidin tetramer, each protein monomer has the potential to specifically bind one biotin-bearing moiety. Herein, by separating various streptavidin species that have had differing numbers of their four potential binding sites blocked, several different types of "linking hub" were obtained, each with a different valency. The identification of these species and the study of the plugging process used to block sites during their preparation were carried out using capillary electrophoresis. Subsequently, a specific species, namely, a trans-divalent linker, in which the two open biotin-binding pockets are approximately opposite one another, was used to concatenate two ∼5 kb pieces of biotin-terminated double-stranded DNA. Following the incubation of this DNA with the prepared linker, a fraction of ∼10 kb strings was identified using gel electrophoresis. Finally, these concatenated DNA strings were stretched in an optical tweezer experiment, demonstrating the potential of the methodology for coupling and extending molecules for use in single-molecule biophysical experiments.
dc.description.confidentialfalse
dc.format.pagination6427-6435
dc.identifier.author-urlhttps://www.ncbi.nlm.nih.gov/pubmed/35224404
dc.identifier.citationMohandas N, Kent LM, Raudsepp A, Jameson GB, Williams MAK. (2022). Progress toward Plug-and-Play Polymer Strings for Optical Tweezers Experiments: Concatenation of DNA Using Streptavidin Linkers.. ACS Omega. 7. 7. (pp. 6427-6435).
dc.identifier.doi10.1021/acsomega.2c00198
dc.identifier.eissn2470-1343
dc.identifier.elements-typejournal-article
dc.identifier.issn2470-1343
dc.identifier.urihttps://mro.massey.ac.nz/handle/10179/68998
dc.languageeng
dc.publisherAmerican Chemical Society
dc.publisher.urihttps://pubs.acs.org/doi/10.1021/acsomega.2c00198
dc.relation.isPartOfACS Omega
dc.rightsCC BY-NC-ND 4.0
dc.rights.urihttps://creativecommons.org/licenses/by-nc-nd/4.0/
dc.titleProgress toward Plug-and-Play Polymer Strings for Optical Tweezers Experiments: Concatenation of DNA Using Streptavidin Linkers
dc.typeJournal article
pubs.elements-id451592
pubs.organisational-groupOther
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