Morgan Brody

2/1/2021

Pd. 11

Section 2.7: DNA replication, transcription, and translation

Semi-conservative replication of DNA

The replication of DNA is semi-conservative and depends on complementary base pairing.

 When a cell prepares to divide two strands of the double helix separate
 New strands are formed by using old strands as a template
 DNA replication is referred to as being semi-conservative
 Hydrogen bonds only form between complementary bases

Obtaining evidence for the theory of semi-conservative replication

Obtaining evidence for scientific theories: Meselson and Stahl obtained evidence for the semi-
conservative replication of DNA.

 Semi-conservative theory of replication needed evidence

 Meselson and Stahl provided strong evidence in elegant experiments:
o They used two different isotopes of Nitrogen like 14N and 15N
o Technique was cesium chloride density gradient centrifugation
o Once the solution is spun in the centrifuge, the greatest density is at the bottom, and
the lowest density is at the top
o They cultured E. coli for 14 generations in a medium where only nitrogen source was
15N
o They then placed it in 14N medium
o They then extracted the DNA and measured it by cesium chloride

Meselson and Stahl’s DNA replication experiments

Analysis of Meselson and Stahl’s results to obtain support for the theory of semi-conservative replication
of DNA.

 Data-based questions from Meselson and Stahl’s experiment

Helicase

Helicase unwinds the double helix and separates the two strands by breaking hydrogen bonds.

 Two strands of DNA must separate so they can become templates

 Separation is carried out by helicase, an enzyme group that uses energy from ATP
  One helicase with six globular polypeptides arranged like a donut, has one strand of DNA go
through the hole and separates the DNA
 DNA can’t be separated while still helical, so helicase causes unwinding of the helix before
separating the strands

DNA polymerase

SNA polymerase links nucleotides together to form a new strand, using the pre-existing strand as a
template.

 The assembly of the new DNA strands is carried out by the enzyme DNA polymerase
 Polymerase always moves along the template strand in the same direction
 It brings nucleotides into the position where the hydrogen bonds could form
 Once the bases are made, it creates covalent bonds between the phosphate group of the free
nucleotide and sugar
 Very few mistakes are made during DNA replication

PCR—the polymerase chain reaction

Use of Taq DNA polymerase to produce multiple copies of DNA rapidly by the polymerase chain reaction
(PCR).

 PCR is a technique used to make many copies of selected DNA sequence

 DNA is loaded into a PCR machine with a cycle of steps repeated
 The machine separates the DNA by heating them to 95 degrees for 15 seconds
 Then quickly cools the DNA to 54 degrees so the strands can re-anneal, but primers are present
 The large excess of primers keeps the DNA from re-annealing, and binds rapidly to target
sequences
 Next phase is synthesis of double-stranded DNA using single strands with primers as templates
 Taq DNA polymerase is used b/c it can resist the brief period at 95 degrees
 Taq DNA polymerase is at its optimum at 72 degrees, at this period it adds about 1,000
nucleotides a minute
 A cycle of PCR can be completed in less than 2 minutes