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Nucleic Acids Research, 1990, Vol. 18, No. 6 1395-1400
© 1990

MOLECULAR BIOLOGY

The basidiomycete Lentinus edodes linear mitochondrial DNA plasmid contains a segment exhibiting a high autonomously replicating sequence activity in Saccharomyces cerevisiae

Yuichi Katayose+, Susumu Kajiwara and Kazuo Shishido*

Department of Life Science, Faculty of Science, Tokyo Institute of Technology Nagatsuta, Midori-ku, Yokohama 227, Japan

*To whom correspondence should be addressed

Received January 3, 1990. Revised February 23, 1990. Accepted February 23, 1990.

A linear DNA plasmid, designated pLLE1, has been isolated from a mitochondrial fraction of a strain of Lentinus edodes. pLLE1 (11.0 kbp) was sensitive to the 3'–5'-acting exonuclease III and resistant to the 5'–3'-acting {lambda} exonuclease. It showed no homology with mitochondrial and nuclear genomic DNAs of plasmidless strain as well as the pLLE1 -harboring host strain of L. edodes. The 1434-bp fragment (sequences) capable of autonomous replication in the yeast Saccharomyces cerevislae (ARS8) was cloned from pLLE1 DNA with Ylp32 (pBR322 containing yeast LEU2 DNA), which displayed a high ARS activity. The cloned 1434-bp fragment was shown to lie near to the end of pLLE1 DNA (nucleotides about 800–2200) and contained three consecutive ARS consensus sequences (A/T)TTTAT(A/G)TTT(A/T) of S. cerevisiae and dispersive eight ARS consensus-like sequences. The subcloned 366-bp fragment containing the three ARSs retained original ARS activity of the 1434-bp fragment.

+Present address Mitsubishi-Kaset Institute of Life Science, Machida, Tokyo, Japan


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