Ovalbumin gene. Action of restriction endonucleases upon DNA coding sequence.

A full length, double-stranded ovalbumin complementary DNA (cDNA) was synthesized in vitro and used to assay the action of 35 restriction endonucleases upon this DNA sequence. Some 22 enzymes, Bal I, BamHl, Blu I and its isoschizomerXho I, Ppu 1,Xba I, Tag I, Sal I, Bgl I, Bgl II, Kpn I, Sst II, Hue II, Hpa I, Hap II, Hha I, Hind11 and its isoschizomer HincII, Hind111 and its isoschizomer Hsu I, EcoRI, and Sma I, failed to cut such a DNA sequence. However, 13 enzymes did cut the DNA. They were Hue III, Pst I and its isoschizomer Xma II, Sst I, Hga I, Xho II, Hph I, EcoRII, HinfI, Mnl I, Mbo I, Mbo II, and ALU I. A detailed map of the restriction enzyme recognition sites for 11 of these restriction enzymes was determined. Since the amino acid sequence of a phosphorylated peptide from hen ovalbumin is known, it was therefore possible to construct a partial nucleotide sequence corresponding to this peptide fragment. Such a sequence could possibly be cut by the three enzymes, EcoRII, HinfI, and Hph I, in close proximity to each other. Since these three enzymes did actually cut the ovalbumin cDNA in such close proximity, we have therefore, tentatively assigned the coding portion of the ovalbumin gene to a region which begins within 200 nucleotides from 5’4erminal end of the gene.