Construction of non-polar mutants in Haemophilus influenzae using FLP recombinase technology
2008

Creating Mutants in Haemophilus influenzae

publication Evidence: high

Author Information

Author(s): Tracy Erin, Ye Fang, Baker Beth D, Munson Robert S Jr

Primary Institution: Center for Microbial Pathogenesis in The Research Institute at Nationwide Children's Hospital, Columbus, OH, USA

Hypothesis

Can we develop a new methodology to construct non-polar mutants in poorly expressed genes of Haemophilus influenzae?

Conclusion

We successfully developed a methodology to create unmarked non-polar mutations in NTHi genes, demonstrating its utility by constructing a non-polar pilA mutation.

Supporting Evidence

  • The methodology allows for the construction of unmarked non-polar mutations in poorly expressed genes.
  • Over 20 mutants have been successfully constructed using this method.
  • The pilABCD operon is essential for type IV pilus biogenesis and transformation in NTHi.

Takeaway

Scientists figured out a new way to change the genes in a germ called Haemophilus influenzae so they can study how it causes sickness. This helps them understand the germ better.

Methodology

The study adapted lambda red/FLP-recombinase-mediated strategies from E. coli for use in NTHi to create non-polar mutations.

Limitations

The methodology may not be suitable for all genes, particularly those that are poorly expressed.

Digital Object Identifier (DOI)

10.1186/1471-2199-9-101

Want to read the original?

Access the complete publication on the publisher's website

View Original Publication