Document Detail

Electrophoretic Heterogeneity Limits the Utility of Streptavidin-β-Galactosidase as a Probe in Free Zone Capillary Electrophoresis Separations.
MedLine Citation:
PMID:  23325029     Owner:  NLM     Status:  Publisher    
Single molecule assays were performed on streptavidin-β-galactosidase using a capillary electrophoresis-based protocol in order to assess the suitability of single molecule β-galactosidase assays for adaptation to the detection of single copies of target DNA. The conjugate was found to have a heterogeneous catalytic rate, showing an average rate of 44,000 ± 24,000 min(-1), which is similar to that of the unmodified enzyme. Electrophoretic mobility was also measured on individual molecules and determined to be -1.32 × 10(-4) ± 0.19 × 10(-4) cm(2)V(-1)s(-1). The variance in mobility was several times that reported for the unmodified enzyme. The electrophoretic heterogeneity was found to result in the formation of a broad window of peaks in the resultant electropherograms of free zone separations of small plugs of streptavidin-β-galactosidase. This range of mobilities largely overlapped with that of the conjugate bound to primer and plasmid containing a target DNA sequence. This overlap suggests that the separation of free conjugate from that bound to target DNA, which is a requirement for application of the single enzyme molecule assay to the detection of target DNA sequences, is not plausible using free zone capillary electrophoresis.
Douglas B Craig; Anna Henderson
Related Documents :
3996319 - Localization of arginine vasopressin-neurophysin ii messenger ribonucleic acid in the h...
4945199 - Isolation of circular deoxyribonucleic acid from salmonella typhosa hybrids obtained fr...
1107199 - Purification of tdna from yeast.
20830429 - Microvalve and micropump controlled shuttle flow microfluidic device for rapid dna hybr...
22961759 - Advances in chromatographic supports for pharmaceutical-grade plasmid dna purification.
2161809 - In situ detection of cytomegalovirus dna in biopsies of aids patients using a hybrido-i...
8104339 - Decatenation activity of topoisomerase iv during oric and pbr322 dna replication in vitro.
10961609 - Incorporation of 3'-azido-3'-deoxythymidine (azt) into fetal dna and fetal tissue distr...
23976919 - Correction: highly mutagenic exocyclic dna adducts are substrates for the human nucleot...
Publication Detail:
Type:  JOURNAL ARTICLE     Date:  2013-1-17
Journal Detail:
Title:  The protein journal     Volume:  -     ISSN:  1875-8355     ISO Abbreviation:  Protein J.     Publication Date:  2013 Jan 
Date Detail:
Created Date:  2013-1-17     Completed Date:  -     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  101212092     Medline TA:  Protein J     Country:  -    
Other Details:
Languages:  ENG     Pagination:  -     Citation Subset:  -    
Chemistry Department, University of Winnipeg, 599 Portage Avenue, Winnipeg, MB, Canada,
Export Citation:
APA/MLA Format     Download EndNote     Download BibTex
MeSH Terms

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

Previous Document:  Recruiting a special sample with sparse resources: Lessons from a study of Gulf War veterans.
Next Document:  The small airways in chronic obstructive pulmonary disease: pathology and effects on disease progres...