The use of primed synthesis by DNA polymerase I to study an intercistronic sequence of phiX-174 DNA.

摘要: A decadeoxynucleotide complementary to ten nucleotides in the major ribosome-protected fragment of phiX-174 plus-strand DNA has been chemically synthesized and used as a primer for polymerase I on template. The sequence first 40 incorporated onto determined. This extends further intercistronic region preceding gene G shows presence another termination codon. was determined by using manganese activating cation which allows ribonucleotides be well deoxyribonucleotides. ribo-substituted product then cleaved specifically at ribonucleotide residues generate series overlapping ribo-terminated fragments whose sequences were sufficient determine complete nucleotides. No evidence misincorporation detected.