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Nucleic Acids Research, 1985, Vol. 13, No. 4 1283-1302
© 1985

Articles

An in vitro system for accurate transcription Initiation of chloroplast protein genes

Emil M. Orozco, Jr.*, John E. Mullet+ and Nam-Hai Chua

Laboratory of Plant Molecular Biology, The Rockefeller University 1230 York Avenue, New York, NY 10021-6399, USA

Received August 10, 1984. Revised January 28, 1985. Accepted January 28, 1985.

We have developed an homologous in vitro system from spinach chloroplasts that correctly initiates transcription of the plastid genes for the large subunit of ribulose-1,5-bisphosphate carboxylase (rbcL) and the ß subunit of the plastid ATPase (atpB) The transcriptionally active extracts from spinach chloroplasts require circular DNA templates for specific initiation. The RNA polymerase activity is insensitive to rifampicin. The extent of transcription in vitro is a function of the extract:template ratio. The efficiency of the rbcL transcription in vitro is more than one transcript per one hundred templates per hour.

*Present address: Agricultural Research Service, U.S. Departments of Agriculture and Departments of Agronomy and Plant Biology, University of Illinois, Urbana, IL 61801, USA

+Present address: Department of Biochemistry and Biophysics, Texas A & M University, College Station, TX 77843, USA


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