BIS 2A Lecture Notes - Lecture 8: Operon, Tryptophan, Repressor

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Variations in the nucleotide sequence of a genome. Can come from errors in replication or from physical damage to dna. Induced: result from exposure to chemicals or environmental agents. Result of random biochemical reactions within the cell. Point: affect a single base pair (usually occur by substitution) Transition substitution: base replaced by a like base. Purine replaced with purine (or pyrimidine w/ pyrimidine) Transversion substitution: base replaced by unlike base. Purine replaced with pyrimidine (or vice versa) Frameshift: insertion or deletion of a nucleotide. Shifts entire sequence of nucleotides from then on. Translocation: gene shifted to a different location. Can be on same or different chromosome. Conservative: similar but not identical amino acids produced. Nonsense: creates stop codon and stops when it shouldn"t. Mutant: organism in which the mutation occurred. Wild type: the norm for a specific trait in the overall population. Ex: most crows are black, that"s the wild type. Mutations: changes from the wild type of a population.

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GENETICS QUESTION: For this experiment we focused on forward genetics: First, his- mutants were identified and then complementation analysis was used in order to determine which of the 7 his genes was mutated. ( The exact data can be seen at the bottom). In this experiment we screened for his- mutants that were generated via a random mutation process resulting from errors in DNA replication or induced by EMS aka Ethyl methanesulfonate. We compared a mutagenized culture that was treated with EMS and an un-mutagenized culture that was not treated with EMS. However, both cultures went trhough an enrichment process to increase the frequency of his- mutants.

Point mutations involve the substitution of one type of base pair for another. If the substitution replaces a purine with a purine (or a pyrimidine with a pyrimidine), it is called a transition. If the substitution replaces a purine with a pyrimidine (or a pyrimidine with a purine), it is called a transversion. Many mutagenic chemicals cause this type of mutation by alkylation of a nucleotide base. The chemically altered base can then be misread during replication or repair. For example the mutagen, ethyl methanesulfonate (EMS), adds an ethyl (-CH2-CH3) group to both guanine or thymine bases causing the modified guanine to pair with thymine and the modified thymine to pair with guanine.

1. Why might one gene be more susceptible to EMS mutagenesis than another gene?

2. Why might we have expected to find a higher frequency of polar mutants (his4A^-B^-C^-) in the unmutagenized culture compared to the mutagenized culture?

Table 2. Experiment 1B class data summary (number of his^-mutants by section) Unmutagenized
Section	1	2	3	*4ABC*	4A	4B	4C	5	6	7	*Total his^-* mutants**
Tuesday	7	1	5	4	2	1	9	2	3	1	35
Wednesday	2	1	14	0	4	2	2	4	8	2	39
Total	9	2	19	4	6	3	11	6	11	3	74
% each mutant	12.1	2.7	25.7	5.4	8.1	4.1	14.9	8.1	14.9	4.1	~100%

Mutagenized
Section	1	2	3	4ABC	4A	4B	4C	5	6	7	*Total his^-* mutants**
Tuesday	2	5	4	2	6	5	5	0	4	1	34
Wednesday	9	1	15	0	9	1	2	2	9	4	52
Total	11	6	19	2	15	6	7	2	13	5	86
% each mutant	12.8	7.0	22.1	2.3	17.4	7.0	8.1	2.3	15.1	5.8	~100%

QUESTION 1

A mutation caused by exposure to gamma rays is called a spontaneous mutation

	a.	True
	b.	False
	c.	Sometimes, depending on the type of base change
	d.	Gamma rays do not cause mutations

QUESTION 2

A recessive mutation affecting an essential biochemical pathway can be detected________

	a.	when the organism is heterozygous for that mutation.
	b.	when the organism is homozygous for that mutation.
	c.	in either homozygous or heterozygous conditions.
	d.	only by sequencing (recessive mutations never show a phenotype).

QUESTION 3

During DNA replication, the newly synthesized chain grows by adding

	a.	nitrogenous bases
	b.	the sugar-phosphate backbone
	c.	nucleosides
	d.	dNTPs

QUESTION 4

During replication, the DNA strand 5â- AAGTCTAGCCTAG -3â will serve as a template for the polymerization of:

	a.	5â- CTAGGCTAGACTT -3â
	b.	5â - TTCAGATCGGATC -3â
	c.	5â - GATCCGATCTGAA -3â
	d.	3â - AAGTCTAGCCTAG -5â

QUESTIOn 5

Given the following DNA sequences, select which statement(s) is(are) correct.

Sequence 1:

5â-TGGACGCTAA-3â

3â-ACCTGCGATT-5â

Sequence 2:

5â-AATCGCAGGT-3â

3â-TTAGCGTCCA-5â

Sequence 3:

5â-ACCTGCGATT-3â

3â-TGGACGCTAA-5â

	a.	Sequences 1 and 2 are the same
	b.	Sequences 1 and 3 are the same
	c.	Sequences 2 and 3 are the same
	d.	Sequences 1, 2 and 3 are all different

QUESTION 6

Repetitive sequences in the genome are hotspots for:

	a.	Deamination
	b.	Depurination
	c.	Thymine dimer formation
	d.	Replication errors

QUESTION 7

Select which statement(s) about the Ames test is(are) correct

	a.	it has been designed to understand mutation repair systems in Salmonella typhimurium
	b.	it uses mammalian liver extract
	c.	it allows to study whether chemical compounds or their enzymatic breakdown products are mutagenic
	d.	it is based on whether a chemical compound causes reversion mutations from his+ to his-
	e.	two of the above are correct
	f.	three of the above are correct

QUESTION 8

Select which statement(s) is(are) correct.

	a.	All DNA strands have a direction, and it is specified by the carbons in the sugar backbone.
	b.	All DNA strands have a direction, and it is specified by hydrogen bonds between nucleotides.
	c.	In order to form proper base pairs in a double stranded DNA molecule, the two strands must run in opposite directions.
	d.	A and C are correct.
	e.	B and C are correct.

QUESTION 9

Studies of gene mutation frequencies have shown that:

	a.	mutations are rare, and genomes are generally stable.
	b.	mutation frequencies differ among organisms and also between genes, suggesting certain genes are more susceptible to mutation.
	c.	mutation frequencies are consistent between organisms, and each region of DNA is equally susceptible to random mutations.
	d.	Both A and B are correct.
	e.	Both A and C are correct.

QUESTION 10

The compound 5-bromodeoxyuridine (BrdU) is a derivative of uracil, and if BrdU becomes incorporated during DNA replication, it pairs with adenine. This compound is best classified as which type of mutagen?

	a.	base analog
	b.	base inducer
	c.	intercalating agent
	d.	oxidative agent
	e.	alkylating agent

QUESTION 11

The rate of mutation of the fruit fly is higher than the rate of mutation of Algae

True

False

QUESTION 12

Thymine dimers are most commonly caused by which of the following?

	a.	X-rays
	b.	Alkylating agents
	c.	U.V. irradiation
	d.	DNA intercalating agents

QUESTION 13

What chemical group is found in the 3â end of a DNA strand?

	a.	an alcohol
	b.	a hydroxyl
	c.	a methyl
	d.	a phosphate

QUESTION 14

What chemical group is found in the 5â end of a DNA strand?

	a.	an alcohol
	b.	a hydroxyl
	c.	a methyl
	d.	a phosphate

QUESTION 15

Which is the correct order of molecules binding to DNA during DNA replication?

	a.	Helicase, SSB, primase, DNA pol III, DNA pol I, ligase
	b.	SSB, DNA pol I, ligase, helicase, DNA pol I, primase
	c.	primase, helicase, DNA pol III, ligase, DNA pol I, SSB
	d.	SSB, helicase, primase, DNA pol III, DNA pol I, ligase

QUESTION 16

Which statement(s) is(are) correct about strand slippage?

	a.	It can cause disorders such as Huntingon disease.
	b.	It is a process that causes mutations altering the number of DNA repeats.
	c.	It is a process that incorporates nucleotide base analogs and trinucleotide repeats.
	d.	a and b are correct.
	e.	b and c are correct.
	f.	a, b and c are correct.

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BIS 2A Lecture Notes - Lecture 8: Operon, Tryptophan, Repressor

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