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

An improved beta-galactosidase alpha-complementation system for molecular cloning in Bacillus subtilis.

P Haima1, D van Sinderen, S Bron

  • 1Department of Microbiology, Center of Biological Sciences, Haren, The Netherlands.

Gene
|September 1, 1990
PubMed
Summary

Researchers optimized the beta-galactosidase alpha-complementation system for Bacillus subtilis cloning. This improved gene expression, stabilized plasmids, and enabled cloning of large DNA fragments up to 29 kb.

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

Relationship between the dietary inflammatory index and immune function during pregnancy - A secondary analysis of the MicrobeMom2 Study.

European journal of clinical nutrition·2025
Same author

Characterization of external cross-talk from silicon photomultipliers in a liquid xenon detector.

The European physical journal. C, Particles and fields·2025
Same author

Highly conserved bifidobacteria in the human gut: Bifidobacterium longum subsp. longum as a potential modulator of elderly innate immunity.

Beneficial microbes·2024
Same author

Measurements of neutrino oscillation parameters from the T2K experiment using <math><mrow><mn>3.6</mn><mo>×</mo><msup><mn>10</mn><mn>21</mn></msup></mrow></math> protons on target.

The European physical journal. C, Particles and fields·2023
Same author

Observation of high-energy neutrinos from the Galactic plane.

Science (New York, N.Y.)·2023
Same author

Evidence for neutrino emission from the nearby active galaxy NGC 1068.

Science (New York, N.Y.)·2022

Area of Science:

  • Molecular Biology
  • Microbiology
  • Biotechnology

Background:

  • The beta-galactosidase alpha-complementation system is crucial for molecular cloning in Bacillus subtilis.
  • Previous methods had limitations in efficiency and stability for large DNA fragment cloning.

Purpose of the Study:

  • To optimize the beta-galactosidase alpha-complementation system for enhanced molecular cloning in Bacillus subtilis.
  • To improve the efficiency and stability of cloning large heterologous DNA fragments.

Main Methods:

  • Optimized translation efficiency of the lacZ delta M15 gene.
  • Segregationally stabilized the plasmid-borne lacZ delta M15 gene via chromosomal integration.
  • Constructed a new lacZ alpha complementing cloning vector with increased unique target sites.

Related Experiment Videos

Main Results:

  • Successfully cloned large heterologous DNA fragments up to at least 29 kb.
  • Demonstrated structural stability of cloned inserts for at least 100 generations.
  • Enhanced cloning vector efficiency and stability in Bacillus subtilis.

Conclusions:

  • The optimized beta-galactosidase alpha-complementation system significantly improves large DNA fragment cloning in Bacillus subtilis.
  • The enhanced system offers greater stability and efficiency for molecular cloning applications.
  • This advancement provides a robust tool for genetic engineering in Bacillus subtilis.