Genetics Ch 6: Dna Replication + Modification

25/10/2010 00:52:00
Linear DNA = multiple replication sites
Many OriRep – about 40,000 bases apart
• Replicated in 15-30 minutes
Replication of full genome (many chromosomes) = 5-10 hours
Okazaki fragments: 1000-2000bp in Prok, 100-200bp in Eukaryotic
Important Replication Proteins
• Gyrase – topoisomerase II (double-stranded break)
• Helicase – Unwinds DNA at replication fork to separate parental
strands
• SSB – stabilizes single-stranded DNA
• Primosome – creates RNA primer
o RNA Primase
• DNA Polymerase – 5’->3’ Polymerization + proofreading functions
In Euk, RNA primer removed by RPA (Replication Protein A)
In Prok, RNA primer removal/replacement by DNA Pol I.
• Both use DNA ligase

Circular DNA can replicate unidirectionally (rolling circle)(plasmids) or

bidirectionally (theta)

Lagging strand synth. leaves overhang once primer removed – telomerase

needed. Telomerase contains built in RNA primer segment (reverse
transcriptase activity)

In PCR, only required enzyme is DNA Polymerase (primers are added)

• Use special high-temperature DNA Pol from bacterial thermophile
TAC Polymerase

Nucleid Acid Hybridization: Denature DNA -> can combine with other
complimentary or almost-complimentary strand.

Restriction Enzymes: Cut duplex DNA at particular sequences

(endonucleases)
 • Often used as genome defense – antivirals. Host DNA is modified
(methylation of C and A), unmodified DNA is vulnerable to
restriction
o Can be symmetric or asymmetric – useful because it leaves
sticky ends.
o Particular RE will chop a genome into a consistent set of
restriction fragments. Different RE = different restriction
map.

Chromosomal Basis of Heredity

Chp 3,2 XX XY
Females = homogametic (XX)
XX XY
Males = heterogametic (XY)
Punnett Square

Birds and butterflies = OPPOSITE, Males = ZZ, Females = ZW

Gregor Mendel – general rules of heredity.

• 1856-1863: pea experiments
• First statistical rules
• Elements of heredity (factors) remain unchanged thru generations
o NOT blending inheritance
• Garden pea: lots of alternative traits, self-fertilization, easy to
artificially fertilize
• Lots of “selfing” = True/pure breeding = P generation
• Monohybrid Cross: cross bw types differing by one trait
•
Round vs Wrinkled
• Determined by one gene
• Encodes starch-branching enzyme 1 (SBE1)
o Synthesizes amylopectic
• Round (W) seeds contain amylopectic – uniform shrinking
 • Wrinkled (w) seeds shrink irregularly (inborn error of starch
metabolism)
• In w (wrinkled): mutation in transposable element interrupts gene
o W smaller than w (transposable element,) can be separated
in a gel
• Homologous genes are “wildtype”
Blocking chlorophyll breakdown = peas stay green longer (yellow are
dominant)
After F1 hybrid self fertilization:
• Green x Yellow = 3 yellow, 1 green (inheritance not sex-linked)
Y

• F2 ratios always like this

Ex: Widow’s Peak: often in 3:1 ratio in humans.

“Particulate” nature of heredity – gamete are equally likel to contain each

half of gene
Self cross of F2:
• ¾ were round  1/3 give only round, 2/3 gave round and wrinkled
in 3:1 ratio
• ¼ were wrinkled  all wrinkled
Test Cross: Unknown genotype crossed w/ homozygous recessive “tester”
• Reveals genotype of unknown – if ½ wrinkled, ½ smooth unknown
= Ww x ww = 2Ww, 2 ww

Dihybrid Cross: cross b/w types differing in 2 characters

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