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Applied and Environmental Microbiology, November 2000, p. 4655-4661, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Isolation and Use of a Homologous Histone H4 Promoter and a Ribosomal DNA Region in a Transformation Vector for the Oil-Producing Fungus Mortierella alpina

Donald A. Mackenzie,* Prasert Wongwathanarat,dagger Andrew T. Carter, and David B. ArcherDagger

Institute of Food Research, Norwich Research Park, Colney, Norwich, Norfolk, NR4 7UA, United Kingdom

Received 6 April 2000/Accepted 29 August 2000

Mortierella alpina was transformed successfully to hygromycin B resistance by using a homologous histone H4 promoter to drive gene expression and a homologous ribosomal DNA region to promote chromosomal integration. This is the first description of transformation in this commercially important oleaginous organism. Two pairs of histone H3 and H4 genes were isolated from this fungus. Each pair consisted of one histone H3 gene and one histone H4 gene, transcribed divergently from an intergenic promoter region. The pairs of encoded histone H3 or H4 proteins were identical in amino acid sequence. At the DNA level, each histone H3 or H4 open reading frame showed 97 to 99% identity to its counterpart but the noncoding regions had little sequence identity. Unlike the histone genes from other filamentous fungi, all four M. alpina genes lacked introns. During normal vegetative growth, transcripts from the two histone H4 genes were produced at approximately the same level, indicating that either histone H4 promoter could be used in transformation vectors. The generation of stable, hygromycin B-resistant transformants required the incorporation of a homologous ribosomal DNA region into the transformation vector to promote chromosomal integration.

* Corresponding author. Mailing address: Institute of Food Research, Norwich Research Park, Colney, Norwich, Norfolk, NR4 7UA, United Kingdom. Phone: 44 1603 255255. Fax: 44 1603 507723. E-mail: donald.mackenzie{at}bbsrc.ac.uk.

dagger Present address: Department of Biotechnology, Faculty of Science and Technology, Thammasat University, Rangsit Center, Patumthanee 12121, Thailand.

Dagger Present address: School of Life and Environmental Sciences, University of Nottingham, University Park, Nottingham NG7 2RD, United Kingdom.

Applied and Environmental Microbiology, November 2000, p. 4655-4661, Vol. 66, No. 11
0099-2240/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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