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A 110 bp palindrome stimulates plasmid integration in yeast

Background and purpose: Perfect inverted repeats, or palindromes, are known as potent inducers of genetic recombination. In our previous work we investigated the influence of a 110 bp palindromic insertion present on a non-replicative plasmid on integration into the yeast genome (19). The palindrome influenced the final outcome of recombination process, but no stimulation or targeting of plasmid integration was observed. In this study we wondered whether plasmid integration would be stimulated when the palindrome was present in both the chromosomal and the plasmid copy of the CYC1 gene. Materials and methods: Yeast strains and plasmids containing palindromic sequence in the CYC1 gene were constructed using conventional methods. Strain construction was followed by Southern blot analysis. Results: When the palindromic sequence was present in both the chromosomal and the plasmid copy of the CYC1 gene the relative efficiency of transformation was increased more then four-fold. During intrachromosomal ("pop-out") recombination the palindrome present in only one copy of the CYC1 gene was lost with very high frequency. Conclusions: Our study may encourage the use of inverted repeats present in the genomes of different organisms for development of palindrome-based integrative vectors.

palindrome ; recombination ; plasmid integration ; yeast

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