Rivista
/
Biologia
/
Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis
JoVE Journal
Biologia
È necessario avere un abbonamento a JoVE per visualizzare questo.  Accedi o inizia la tua prova gratuita.
JoVE Journal Biologia
Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis

Generation of Enterobacter sp. YSU Auxotrophs Using Transposon Mutagenesis

DOI:
10.3791/51934-v

•

13:31 min

•

October 31, 2014

•

1Department of Biological Sciences, Center for Applied Chemical Biology,Youngstown State University

Capitoli

  • 00:05Titolo
  • 02:27Electroporation of Competent Cells
  • 04:35Gridding Transformants and Replica Plating to Determine the Auxotroph Phenotype
  • 08:02Gene Rescue
  • 11:13Results: Generation of Auxotrophic Transformants in Enterobacter sp. YSU
  • 12:39Conclusion

Summary

Traduzione automatica

Traduzione automatica

Enterobacter sp. YSU grows in glucose minimal salts medium. Auxotrophs are generated by transforming it with a transposome which randomly inserts itself into the host genome. Mutants are found by replica plating from complex medium to minimal medium. Interrupted genes are identified by gene rescue and sequencing.

Tags

Enterobacter Sp. YSUAuxotrophsTransposon MutagenesisTn5 TransposomeM-9 Minimal MediumR6K Gamma Replication OriginKanamycin ResistanceSanger SequencingBLASTGene Identification

Article

Embed

AGGIUNGI ALLA PLAYLIST

Usage Statistics

Video correlati

Generation of Stable Transgenic C. elegans Using Microinjection

Homemade Site Directed Mutagenesis of Whole Plasmids

Generation of RNA/DNA Hybrids in Genomic DNA by Transformation using RNA-containing Oligonucleotides

Mutagenesis and Functional Analysis of Ion Channels Heterologously Expressed in Mammalian Cells

An Introduction to Worm Lab: from Culturing Worms to Mutagenesis

Mutagenesis and Functional Selection Protocols for Directed Evolution of Proteins in E. coli

Single Oocyte Bisulfite Mutagenesis

Rapid and Efficient Zebrafish Genotyping Using PCR with High-resolution Melt Analysis

Efficient iPS Cell Generation from Blood Using Episomes and HDAC Inhibitors

A Protocol for Functional Assessment of Whole-Protein Saturation Mutagenesis Libraries Utilizing High-Throughput Sequencing

Read Article