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

A dual protein expression system in Bacillus subtilis.

Annie Y Chan1, Mei M Chan, Hei M Lo

  • 1Department of Zoology, The University of Hong Kong, Pokfulam Road, Hong Kong, China.

Protein Expression and Purification
|December 4, 2002
PubMed
Summary

This study presents a dual expression system for Bacillus subtilis, enabling simultaneous overexpression of two proteins. Gene position relative to the promoter influenced expression levels, with tandem gene duplication not increasing production.

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

Expression of Concern: The bi-directional translocation of MARCKS between membrane and cytosol regulates integrin-mediated muscle cell spreading.

Journal of cell science·2023
Same author

Modulating the activities of chloroplasts and mitochondria promotes adenosine triphosphate production and plant growth.

Quantitative plant biology·2023
Same author

What is quantitative plant biology?

Quantitative plant biology·2023
Same author

ATP translocation and chloroplast biology.

National science review·2021
Same author

Multiple Kinases Can Phosphorylate the N-Terminal Sequences of Mitochondrial Proteins in <i>Arabidopsis thaliana</i>.

Frontiers in plant science·2018
Same author

FdC1 and Leaf-Type Ferredoxins Channel Electrons From Photosystem I to Different Downstream Electron Acceptors.

Frontiers in plant science·2018

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Microbial Genetics

Background:

  • Simultaneous overexpression of multiple proteins is crucial for various biotechnological applications.
  • Bacillus subtilis is a key host for industrial enzyme production.

Purpose of the Study:

  • To develop and characterize a dual expression system for simultaneous overexpression of two genes in Bacillus subtilis.
  • To investigate factors affecting gene product yields in a tandem expression construct.

Main Methods:

  • Engineered two B. subtilis genes (xylanase and glucanase) with independent Shine-Dalgarno sequences.
  • Cloned genes in tandem into a transfer vector for transformation into B. subtilis.
  • Utilized a bacteriophage phi105 promoter for thermally induced transcription.

Related Experiment Videos

Main Results:

  • Expression levels correlated with gene position relative to the phi105 promoter, regardless of gene identity.
  • The second gene insert exhibited lower expression, potentially due to resource limitation, mRNA instability, or transcriptional termination.
  • Tandem gene duplication did not enhance overall protein production.

Conclusions:

  • The developed dual expression system allows for simultaneous protein overexpression in B. subtilis.
  • Positional effects significantly impact gene expression in tandem constructs.
  • Further optimization is needed to overcome limitations in second-gene expression for enhanced yields.