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Generation of a Gene-disrupted Streptococcus mutans Strain Without Gene Cloning
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JoVE Journal Genetics
Generation of a Gene-disrupted Streptococcus mutans Strain Without Gene Cloning

Generation of a Gene-disrupted Streptococcus mutans Strain Without Gene Cloning

DOI:
10.3791/56319-v

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12:06 min

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October 23, 2017

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, , ,
1Department of Translational Research,Tsurumi University School of Dental Medicine, 2Endowed Department of International Oral Health Science (affiliated with Department of Translational Research),Tsurumi University School of Dental Medicine, 3Cariology and Operative Dentistry, Graduate School of Medical and Dental Sciences,Tokyo Medical and Dental University

Chapters

  • 00:05Title
  • 01:14Genomic DNA Extractions from S. mutans
  • 04:01PCR Amplification
  • 06:43Cell Transformation
  • 07:43Verification of Gene Disruption
  • 08:41Phenotypic Analysis
  • 10:16Results: 2-Step Fusion PCR Generates S. mutans Strains with Gene Disruptions
  • 11:26Conclusion

Summary

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We describe a facile, rapid, and relatively inexpensive method for the generation of gene-disrupted Streptococcus mutans strains; this technique may be adapted for the generation of gene-disrupted strains of various species.

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Keywords: Streptococcus MutansGene DisruptionTwo-step Fusion PCRElectroporationNested PrimersGenomic DNA ExtractionBead-beating

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