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Nucleic Acids Research, 1988, Vol. 16, No. 14 6265-6278
© 1988

Articles

Escherichia coli RNase D: sequencing of the rnd structural gene and purification of the overexpressed protein

Jiren Zhang and Murray P. Deutscher

Department of Biochemistry, University of Connecticut Health Center Farmington, CT 06032, USA

Received February 1, 1988. We have determined the nucleotide sequence of a 1.4-kb-pair fragment of the E. coli chromosome that carries the complete rnd gene encoding RNase D, a putative tRNA processing enzyme. The coding region of rnd extends for a total of 1128 nucleotides beginning at an initiator UUG codon and terminating at a UAA codon, and encodes a 375-amino acid polypeptide of 42,679 daltons, consistent with the known size of RNase D. A rapid purification procedure was developed for isolation of RNase D from strains overexpressing the enzyme. The N-terminal sequence and the amino acid composition of the homogenous protein were in excellent agreement with those derived from the sequence of the rnd gene.


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