Denatured toops (single-stranded Denaturation mapp ng the denatured loops are reproducible and thus can be used as points of reference DNA replication starts at specific- origins)
Denatured loops (single-stranded) Nondenatured DNA (double-stranded) Denaturation mapping: the denatured loops are reproducible and thus can be used as points of reference (DNA replication starts at specific origins)
6. The chemistry of DNA polymerization was revealed by in vitro studies using a DNA polymerase purified from E coli Arthur Kornberg purified DNA polymerase I,a monomeric 103 kDa protein, from E coli in 1955 and revealed the major features of the dna synthesis process DNA polymerase I was found to catalyze DNA polymerization in vitro in the presence of a single stranded DNA template, a preexisting primer with a free 3-OH group and the dNTPs(using radioactive precursor nucleotides to label and trichloroacetic acid to precipitate the newly synthesized DNA)
6. The chemistry of DNA polymerization was revealed by in vitro studies using a DNA polymerase purified from E. coli • Arthur Kornberg purified DNA polymerase I, a monomeric 103 kDa protein, from E. coli in 1955 and revealed the major features of the DNA synthesis process. • DNA polymerase I was found to catalyze DNA polymerization in vitro in the presence of a singlestranded DNA template, a preexisting primer with a free 3`-OH group and the dNTPs (using radioactive precursor nucleotides to label and trichloroacetic acid to precipitate the newly synthesized DNA)
The fundamental reaction for dNa synthesis is a nucleophilic attack by the 3-OH group of the growing strand on the 5-a-phosphorus of a incoming dNTP selected via base-pairing: the newly synthesized dna is al ways extended in the 5 to 3 direction
• The fundamental reaction for DNA synthesis is a nucleophilic attack by the 3`-OH group of the growing strand on the 5`-a-phosphorus of a incoming dNTP selected via base-pairing: the newly synthesized DNA is always extended in the 5` to 3` direction
Incoming O deoxynucleoside 5-triphosphate O-P-O O=P-0 0=P-O O=P-0 5 Growing O=P-0 OH OH DNA strand OH primer G T Template DNA strand 35 De
H H-N A H N G N-HIIIN 心N CH3OH—N N N一H N H TN—HNA H N-H CNH—N OuH-NA N H N一H1O CH. O C NHNG-N TN一H10 N N OIH-N G OIIH-N H-N H (a)