fulltext.study @t Gmail

Tools for advanced and targeted genetic manipulation of the β-lactam antibiotic producer Acremonium chrysogenum

Paper ID Volume ID Publish Year Pages File Format Full-Text
23333 43433 2014 12 PDF Available
Title
Tools for advanced and targeted genetic manipulation of the β-lactam antibiotic producer Acremonium chrysogenum
Abstract

•We developed several molecular genetic tools for Acremonium chrysogenum.•A fungal recipient for homologous recombination was generated.•An inducible promoter was established.•The FLP/FRT recombination was applied to Acremonium chrysogenum.•Marker recycling resulted in a fungal strain, devoid of foreign genes.

Acremonium chrysogenum is the major producer of the β-lactam antibiotic cephalosporin C and therefore of great importance for the pharmaceutical industry. However, this filamentous fungus is known to reproduce solely by asexual means, shows only sporadic conidiospore production, and has gradual fragmentation of the vegetative mycelium into arthrospores. Due to these peculiar growth characteristics and life style, strain improvement by recombinant technologies is much more challenging than for other biotechnologically relevant fungi. Here, we describe several molecular tools for genetic engineering of A. chrysogenum, including a ΔAcku70 deletion strain for homologous recombination. No physiological or morphological changes occurred due to deletion of the ku70 gene or integration of the nat1 cassette in this recipient strain. We also used a xylose-inducible promoter from Sordaria macrospora (Smxyl) to demonstrate induction of the gfp reporter gene in A. chrysogenum. The Smxyl promoter was used for construction of a vector molecule to develop a one-step FLP/FRT recombination system in A. chrysogenum. This system was then used in the ΔAcku70 deletion strain to construct a marker-free recipient strain for targeted DNA insertion into genomic DNA. The applicability of our tools was demonstrated by construction of a marker-free transgenic strain, lacking any foreign genes.

Keywords
Acremonium chrysogenum; Cephalosporin C; FLP/FRT recombination system; Inducible promoter; Targeted insertion of foreign DNA
First Page Preview
Tools for advanced and targeted genetic manipulation of the β-lactam antibiotic producer Acremonium chrysogenum
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 169, 10 January 2014, Pages 51–62
Authors
, , , , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us