Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Chitobiase, a new reporter enzyme

D Y Kalabat1, J M Froelich, T K Phuong

  • 1San Diego State University, CA, USA.

Biotechniques
|December 24, 1998
PubMed
Summary

N,N'-diacetylchitobiase (chitobiase) from Vibrio harveyi is a stable reporter enzyme for gene expression studies. Its use in bacteria lacking native enzymes allows for effective monitoring of gene fusions.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Controlled dissolution of freeform 3D printed carbohydrate glass scaffolds in hydrogels using a hydrophobic spray coating.

Additive manufacturing·2019
Same author

Metal-activated C-peptide facilitates glucose clearance and the release of a nitric oxide stimulus via the GLUT1 transporter.

Diabetologia·2007
Same author

Generation of retroviral packaging and producer cell lines for large-scale vector production and clinical application: improved safety and high titer.

Molecular therapy : the journal of the American Society of Gene Therapy·2000
Same author

Role of DNA methylation at GATC sites in the dnaA promoter, dnaAp2.

Journal of molecular microbiology and biotechnology·2000
Same author

Chromosomal insertions localized around oriC affect the cell cycle in Escherichia coli.

Biochimie·1999
Same author

HMO productivity: back to basics.

Managed care interface·1997

Area of Science:

  • Microbiology
  • Molecular Biology
  • Biochemistry

Background:

  • N,N'-diacetylchitobiase (chitobiase) is a reporter enzyme derived from Vibrio harveyi.
  • It offers high stability and specific substrate hydrolysis.
  • Its absence in E. coli and availability of substrates make it advantageous.

Purpose of the Study:

  • To develop and characterize chitobiase as a reporter gene for studying gene expression.
  • To construct plasmid vectors for in-frame gene fusions with chitobiase.
  • To analyze gene regulation using single-copy chromosomal fusions.

Main Methods:

  • Construction of plasmid vectors with multiple cloning sites for chitobiase fusions.
  • Integration of gene fusions into the bacterial chromosome via site-specific recombination.
  • Assay of chitobiase activity in single-copy chromosomal fusions.

Main Results:

  • Plasmid vectors were successfully constructed for in-frame fusions to chitobiase.
  • Site-specific recombination enabled single-copy chromosomal integration of dnaA-chb fusions.
  • Chitobiase activity was used to examine the effects of dnaA regulatory region deletions.

Conclusions:

  • Chitobiase is a versatile and reliable reporter enzyme for gene expression studies in bacteria.
  • The developed system allows for precise analysis of gene regulation at the single-copy level.
  • This approach is applicable to bacteria lacking endogenous N-acetyl-beta-D-glucosaminidase activity.

Related Experiment Videos