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Nucleic Acids Research, 1995, Vol. 23, No. 10 1704-1709
© 1995

MOLECULAR BIOLOGY

The processing of macronuclear-destined DNA sequences microinjected into the macronuclear anlagen of the hypotrichous ciliate Stylonchia lemnae

Jian-Ping Wen, Claudine Eder1 and Hans J. Lipps*

Institut für Zellbiologie, Universität Wrtten/Herdecke Stockumer Strasse 10, 58448 Witten, Germany 1Max Delbrück Centrum für Molekulare Medizin Berlin, Germany

* To whom correspondence should be addressed

Received February 10, 1995. Revised March 30, 1995. Accepted March 30, 1995.

We describe the construction of a vector carrying the micronuclear versions of two macronuclear DNA molecules, one of which was modified by the insertion of a polylinker sequence. This vector was injected into the polytene chromosomes of the developing ma-cronucleus of Stylonychla and its processing during further macronuclear development and Its fate in the mature macronucleus were analyzed. In up to 30% of Injected cells the modified macronuclear DNA sequence could be detected. While the Internal eliminated sequences (IES) present in the macronuclear precursor DNA sequence are still retained in the mature macronucleus, the modified macronuclear DNA sequence Is correctly cut out from the vector, telomeres are added de novo and it Is stably retained In the macronucleus during vegetative growth of the cells. This vector system represents an experimental system that allows the identification of DNA sequences involved in the processing of macronuclear DNA sequences during macronuclear development.


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