Purification of nucleotide sequences suitable for expression in

Details Purification of nucleotide sequences suitable for expression in Purification of nucleotide sequences suitable for expression in

1979-11-23

1981-08-11

Warden, Robert J.

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving nucleic acid

435 89, 435 91, C12Q 168

Patent

active

042834899

ABSTRACT:
A method has been discovered for purifying a specific desired DNA sequence, starting from RNA heterogeneous in length and sequence. The steps of the method include making complementary DNA transcripts of the RNA by means of an enzyme such as reverse transcriptase, subjecting the DNA transcripts to the action of one or more selected restriction endonuclease enzymes, and fractionating the fragments produced by endonuclease action according to their length. By this method it is possible to isolate homogeneous length DNA fragments complementary to RNA sequences present in the original preparation in as low a frequency as two percent. A method is also disclosed for further purifying the homogeneous length fragments and for determining their final purity. Using the disclosed methods, a DNA fragment approximately 550 nucleotides in length coding for a portion of the peptide hormone, human chorionic somatomammotropin, has been purified to greater than 99% purity.

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