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Nucleic Acids Research, 1995, Vol. 23, No. 13 2526-2530
© 1995


MOLECULAR BIOLOGY

Disruption of the Crithidia fasciculata RNH1 gene results in the loss of two active forms of ribonuclease H

Dan S. Ray* and Jane C. Hines

Molecular Biology Institute and Department of Biology, University of California Los Angeles, CA90095-1570, USA

*To whom correspondence should be addressed

Received March 2, 1995. Accepted May 25, 1995.

Both prokaryotic and eukaryotic cells contain multiple forms of ribonuclease H, a ribonuclease that specifically degrades the RNA strand of RNA-DNA hybrids and which has been implicated in the processing of Initiator RNAs and In the removal of RNA primers from Okazaki fragments. The Crithidia fasciculata RNH1 gene encodes an RNase H and was shown to be a single-copy gene in this diplold trypanosomatld. The RNH1 gene has been disrupted by targeted gene disruption using hygromycin or G418 drug-resistance cassettes. Major active forms of RNase H (38 and 45 kDa) were observed on activity gels of extracts of wild-type cells or cells in which one allele of RNH1 was disrupted. Both the 38 and 45 kDa activities were absent In extracts of cells In which both alleles of RNH1 were disrupted indicating that both forms of the C.fasclculata RNase H are encoded by the RNH1 gene.


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