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AUD4, a new amplifiable element from Streptomyces lividans

Institute of Industrial Genetics, University of Stuttgart, Allmandring 31, 70569 Stuttgart, Germany¹

Author for correspondence: Josef Altenbuchner. Tel: +49 711 685 7591. Fax: +49 711 685 6973. e-mail: joe{at}gensun.biologie.uni-stuttgart.de

After transformation of the Streptomyces lividans chloramphenicol-sensitive, arginine-auxotrophic mutant strain AJ100 with a derivative of plasmid SCP2, some of the regenerated protoplasts contained an 8·2 kb DNA sequence amplified to several hundred copies per chromosome. The corresponding non-amplified sequence, called AUD4, was isolated from a phage genomic library of S. lividans 1326. Two cytosine residues were the only directly repeated nucleotides at the ends of the element, indicating that AUD4 is a class I amplifiable sequence. The element mapped in the AseI-D fragment of the S. lividans chromosome, where other class I amplifications have been described. The complete element was sequenced and 10 ORFs were identified. Some of the deduced proteins are highly conserved in other organisms but a putative function could be attributed to only a few of them. Duplication of AUD4 by integration of an Escherichia coli plasmid carrying various parts of AUD4 and a thiostrepton-resistance gene in S. lividans AJ100, ZX7 or TK64 induced amplification of the integrated plasmid, AUD4 or both at high frequency.

Keywords: DNA amplification, deletion, genetic instability, class I amplifiable elements

Abbreviations: ADS, amplified DNA sequence; AUD, amplifiable unit of DNA; HT, Hickey–Tresner

The GenBank accession number for the sequence reported in this paper is AF072709.

^a Present address: Cardiovascular Biology, Pfizer Central Research, Pfizer Limited, Sandwich, Kent CT13 9NJ, UK.

^b Present address: Klinische Chemie, Franz Josef Strauß-Allee 11, 93053 Regensburg, Germany.

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