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Nucleic Acids Research, 1985, Vol. 13, No. 17 6237-6248
© 1985

Articles

The ribosomal RNA promoter of Acanthamoeba castellanii determined by transcription in a cellfree system

Preecha Kownin, Calvin T. lida, Sheryl Brown-Shimer and Marvin R. Paule

Department of Biochemistry, Colorado State University Fort Collins, CO 80523, USA

Received October 24, 1984. Revised February 2, 1985. Accepted August 6, 1985.

The DNA sequences required for faithful initiation of ribosomal RNA transcription were determined. BAL–31 digestion was used to modify the rDNA template by introducing deletions from its 3'–and 5'– ends. The resulting mutant DNAs were tested for template activity individually or in competition with wild type utilizing an in vitro transcription system from Acanthamoeba castellanii. The results identify the sequence extending from –31 to +8 to be absolutely required for transcription. In addition; when the region between –47 and –32 is left intact, transcription is augmented.


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