Genetics Final Exam

• A

• C

B

• Transition at 11: Met Thr Gly Ser Gln Leu Tyr

• Transition at 13: Met Thr Gly Asn

• Deletion at 7: Met Thr Ala Ile Asn Tyr Ile

• Transversion at 15: Met Thr Gly Asn His Leu Tyr

• Addition after 6: Met Thr Thr Gly Asn Gln Leu Tyr

• Transition at 9: Met Thr Gly Asn Gln Leu Tyr

A

D

• GUA, GUG

• B

nonsense, frameshift, insertion

D
Ctc—> Ct/Cc→ Cc

A,D

Class I:

• also called transposons

• transpose through an RNA intermediates

• more common in euks than other types of transposons

• present in euks & absent from bac

• inserted into genome by reverse transcriptase

Class II:

• also called DNA transposons

• transpose as DNA

• present in bac & euks

• may transpose thru replicative or non-replicative transposition

BOTH:

• flanking direct repeats are at either end of the transposable element

• can potentially be inserted at any location in the genome with a recognized cutting sequence

• also called jumping genes

B

Orthologs:

• A1 & B1

• A1 & B2

• A1 in species 1 & A1 in species 2

• A1 in species 2 & A2 in species 3

• All copies of A2

Paralogs:

• B1 & B2

• A1 & A2 in species 2

Comparative:

• analyzes multiple genomes to determine gene function

• examines evolutionary relationships btwn organisms

Functional:

• investigates cellular roles of genes & gene products

• focuses on transcription & translation

Structural:

• focus on determining nucleotide sequence of a genome

• concerns physical organization of DNA

B

A,D,E

Pro:

• relatively small genome

• 500-7000 genes

• ~1 gene/1000 bps

• few, if any, introns

Euk:

• relatively large, variable size genome

• 6000-30000 genes

• ~1 gene/2000-100000 bps

• multiple introns

D,E

Denisovan

5’ end

5’-CTATCAAAAGCAT-3’

5’-ATCAAAAGC-3’

5’-TCAAAAGCATAGAGGTACC-3’

5’-AAAAGCATAGAGGTACC-3’

5’GAGGTACCCTTC-3’

C

• create a cloning vector that contains an origin of replication recognized by e coli, an antibiotic resistance gene, and one or more restriction sites

• cut a solution of double-stranded DNA containing the gene encoding insulin and the cloning vector with the same restriction enzyme and mix them together

• add a solution of plasmid to a culture of e coli

• screen e coli for recombinant plasmids by adding antibiotic to the culture and selecting only the colonies that survive

disrupt target gene by inserting neo+ and link tk+ to disrupted gene

• transfer disrupted gene into stem cells from a black mouse

• allow recombination with genome to generate neo+ tk- cells

• select for neo+ and select against tk+ to isolate cells from homologous recombination

• inject neo+ cells into white early embryo and implant into surrogate

• identify chimeric progeny with target-gene knockout cells

breed mice to generate homozygous knockouts

• restriction enzyme

• restriction site

• dna ligase

• ligation

• recombinant plasmid

• C

• A

• B

• E

• D

AAATTACAGAATCACATA

D

AGC CATAGCA

• A

• A,C

BACs:

• transformed as a circular chromosome

• contains par genes originating from the F’ plasmid

YACs:

• contains a centromere sequence

• transformed as a linear chromosome

• contains a telomere sequence

Both:

• used for cloning of long (>10 kbp) DNA fragments

• contains an origin of replication

B,C,D

• 2

• 8

• ~1 billion

• A

• denature with heat

• add primers

• add DNA poly and deoxynucleotides

• Determine the DNA sequence of the region of the genome containing PDK-4

• Identify a protospacer-adjacent motif (PAM) adjacent to PDK-4 in the genome

• Generate Cas9 mRNA and sgRNA specific to the target sequence using in vitro transcription

• Isolate single cell mouse embryos and inject them with Cas9 mRNA and sgRNA

DNA to be cloned

• mature mammalian mRNA is isolated

• reverse transcriptase, provided with a primer, synthesizes a strand of cDNA without introns

• the mRNA is hydrolyzed by raising the pH

• a primer site is created in the single-stranded DNA

• a complementary DNA strand is synthesized, forming a double0stranded cDNA molc

Double-stranded DNA

D

A

A

D

Epi:

• a pt has angelman syndrome because his mother has the syndrome

• DNA methylation decreases as cells age

Environmental:

• an essential gene in the fruit fly is inactive at 29ºC and active at 22ºC

• matt Is taller than his dad because matt eats a more nutritious diet

Both:

• the offspring of a mouse are anxious because she experienced stress while pregnant

• chris’ high sugar diet causes a demethylation of genes associated with type II diabetes

Neither:

• a newborn is diagnosed with Down syndrome even though it does not run in the family

• C

• B

• prior to replication, CpGs are fully methylated

• the replicated DNA strand is synthesized unmethylated

• methyltransferase binds to DNA methyl groups

• any unmethylated DNA is methylated

• fully methylated DNA is produced

A,C,D

D

• crosslink DNA binding proteins to DNA

• shear protein bound DNA into fragments

• immunoprecipitate target protein

• reverse crosslinking of DNA and protein

• purify and sequence DNA

• A