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

Nucleic Acids Res.1988 Jul 25;16(14A):6265-78. doi:10.1093/nar/16.14.6265
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摘要


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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