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Genetics Ch 6DNAREP

Genetics Ch 6DNAREP

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Published by: April Iman on Oct 25, 2010
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Genetics Ch 6: Dna Replication + Modification1/8/2010 1:16:00 PM

Linear DNA = multiple replication sites Many OriRep ± about 40,000 bases apart y 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 y Gyrase ± topoisomerase II (double-stranded break) y Helicase ± Unwinds DNA at replication fork to separate parental strands y SSB ± stabilizes single-stranded DNA y Primosome ± creates RNA primer o RNA Primase y 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. y 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) y 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) y Often used as genome defense ± antivirals. Host DNA is modified (methylation of C and A), unmodified DNA is vulnerable to restriction

o Particular RE will chop a genome into a consistent set of restriction fragments.o Can be symmetric or asymmetric ± useful because it leaves sticky ends. Different RE = different restriction map. XX XY XX XY .

self-fertilization. 1 green (inheritance not sex-linked) .2 Females = homogametic (XX) Males = heterogametic (XY) Punnett Square Birds and butterflies = OPPOSITE. easy to artificially fertilize y Lots of ³selfing´ = True/pure breeding = P generation y Monohybrid Cross: cross bw types differing by one trait y Round vs Wrinkled y Determined by one gene y Encodes starch-branching enzyme 1 (SBE1) o Synthesizes amylopectic y Round (W) seeds contain amylopectic±uniform shrinking y Wrinkled (w) seeds shrink irregularly (inborn error of starch metabolism) y In w (wrinkled): mutation in transposable element interrupts gene o W smaller than w (transposable element. Females = ZW Gregor Mendel ± general rules of heredity. Males = ZZ.) can be separated in a gel y Homologous genes are ³wildtype´ Blocking chlorophyll breakdown = peas stay green longer (yellow are dominant) After F1 hybrid self fertilization: y Green x Yellow = 3 yellow. y 1856-1863: pea experiments y First statistical rules y Elements of heredity (factors) remain unchanged thru generations o NOT blending inheritance y Garden pea: lots of alternative traits.Chromosomal Basis of Heredity Chp 3.

2 ww Dihybrid Cross: cross b/w types differing in 2 characters .YY Yy Yy yy y F2 ratios always like this Ex: Widow¶s Peak: often in 3:1 ratio in humans. ³Particulate´ nature of heredity ± gamete are equally likel to contain each half of gene Self cross of F2: y ¾ were round 1/3 give only round. ½ smooth unknown = Ww x ww = 2Ww. 2/3 gave round and wrinkled y in 3:1 ratio ¼ were wrinkled all wrinkled Test Cross: Unknown genotype crossed w/ homozygous recessive ³tester´ y Reveals genotype of unknown ± if ½ wrinkled.

1/8/2010 1:16:00 PM .

1/8/2010 1:16:00 PM .

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