BIO 2306 Lecture Notes - Lecture 25: Frameshift Mutation, Sickle-Cell Disease, Replica Plating

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28 Aug 2016

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Genetic mutations: cis-trans test, cross hetero and get a few wild types then they are different alleles, complementation. If there is then they are different genes and if not, then they are the same gene: types of mutations, base substitution. Transition: subs of a purine for a purine or an pyr for pyr. Transversion: subs a purine for a pyr or a pyr for purine, subs an a for a c or an a for a t, sickle cell anemia is caused by this. Difference in one amino acid: frame shift mutations. Base insertion: every group of three bases will be different after the insertion point, the protein is vastly effected. Base deletion: same as above, pre-adaptive v post-adaptive mutations, pre-adaptive. In a colony of bacteria a few of them will have alleles that have resistance for something. The something did not cause the mutation it already existed: post-adaptive, mutations can after exposure, lederbergs replica plating.

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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%

1. An occurrence of a gene made larger by trinucleotide repeats is:

Allelic expansion

Nucleotide expansion

Translocation mutation

Transformation

2. a chemical that can damage and/or change DNA is called a/an:

Allele

Endonuclease

Vector

Mutagen

3. An occurrence when a section of a chromosome relocates itself to an entirely different (non-homologous) chromosome is called a/an:

Inversion mutation

Translocation mutation

Transformation mutation

Duplication mutation

4. The tandem repeat in the sequence GGGAAGGGAAGGGAAGGGAAGGGAAG is:

GGA

GGGAA

GGAAG

GGAAGGG

A disease characterized by abnormally shaped hemoglobin is called:

Cystic Fibrosis

Sickle Cell Anemia

Marfan Syndrome

Leukemia

5. A point mutation that causes a substitution of a stop codon with an amino acid and leads to the formation of a longer protein is a:

Nonsense mutation

Missense mutation

Sense mutation

Frameshift mutation

6. Addition or deletion of nucleotides in a DNA sequence is known as a:

Nonsense mutation

Missense mutation

Sense mutation

Frameshift mutation

7. The least severe type of chromosomal mutation is:

Point

Frameshift

Inversion

Translocation

8. Chemicals inserting themselves into DNA can cause a:

Missense mutation

Nonsense mutation

Sense mutation

Frameshift mutation

9. A genetic condition caused by allelic expansion is:

Familial hypercholesterolemia

Fragile X syndrome

Alkaptonuria

Galactosemia

10. Which of the following is most likely the original DNA strand if the mutated DNA strand is ATAGUUGATGUA ?

ATAGAAGATGAA

ATAGCCGATGCA

ATAGGGGATGGA

ATAGTTGATGTA

11. An unbalanced chromosomal mutation would include a/an:

duplication (insertion)

inversion

translocation

denaturation

12. Many translocation mutations are found to be involved with:

cancers

sickle cell anemia

Huntington disease

cystic fibrosis

13. A common repeat throughout the human genome that is approximately 300 bases in length is called a/an:

EcoRI repeat

Hind repeat

Exo repeat

Alu repeat

14. A balanced chromosomal mutation includes:

imprintation

denaturation

inversion

deletion

salmondinosaur263

1. In Humans, many types of cancers have been linked to?

A. Duplications
B. Inversions
C. translocations
D. deletions
E all of these

2. 5- bromoouracil is mutagenic because it is a(n)

a. base-modifying agent
b. base analog
c. intercalating agent
d. alkylating agent.

3.Ultraviolet radiation creates __________ in DNA

A. 5-methyl cytosine residues
B. breakages
C point mutations
D. mutational hot spots
E thymine dimers

4. The telomeres of chromosomes is composed of

A. heterochromatin
b. nucleosome
c. histone
d. achromatic

5. In eukaryotes, a replicon is

a. the distance between the origin of replication and thetermination of replication
b. a single chromosome
c. the entire genome
d. one DNA strand of a single chromosome

6. In a nonreciprocal interchomosomal translocation, a segment of achromosome is

a. exchanged with an identical segment on a differentchromosome
b. exchanged with an identical segment on a sister chromatid
c. transferred to a different chromosome without a reciprocalexchange of genetic material
d. transferred to a new location on the same chromosome without areciprocal exchange of genetic material

7. Somatic mutations are always.

a. spontaneous
b. carcinogenic
c. heritable
d. lethal
e. none of these

8. A woman with an XO genotype is

a. aneuploid
b. euploid
c. diploid
d. poluploid

9. Spontaneous mutations may occur as a result of

a. all of these
b. errors in DNA replication
c. transposition
d. none of these
e. normal cell biochemistry

10. In the semi conservative model of DNA replication, progenydouble helices consist of

a. one parental DNA strand and one new strand
b. one parental DNA strand and one new RNA strand
c. two new RNA strands
d. two parental DNA strands
e. two new DNA strands

11. A mutation in a gene may alter a cellâs

a. phenotype
b. genotype and phenotype
c. genotype

12. During replication, the direction of synthesis of new DNA fromthe leading strand is

a. both 5â to 3â and 3â to 5â
b. none of these
c. 3â to 5â
d. from left to right only
e. 5â to 3â only

13. A mutation that results in a switch from a purine-pyramidingbase pair to a pyramiding-purine base pair is a _________mutation.

a. neutral
b. transition
c. missense
d. transversion
e. nonsense

14. A trisomic human cell would contain how many chromosomes?

a. 45
b. 46,
c. 47.
d. 48

15. In a strand of DNA, a _____________ bond connects the phosphategroup of one nucleotide to the sugar of the adjacentnucleotide

a. hydrogen
b. ester
c. phosphodiester
d. phosphate
e. disulfide

16. Cri-du-chat syndrome is caused by which type of chromosomalmutation?

a. deletion
b. duplication
c. translocation
d. transversion
e. inversion

17. During replication, the role of DNA primase is to

a. add nucleotide to the growing DNA chain
b. synthesize a short chain for RNA
c. untwist the double helix at the origin of replication
d. relax the super coiling of the double helix
e. synthesize the primo some

18. A(n) ____________ inversion involves the Centro mere

a. chromocentric
b. paracentric
c. epicentric
d. percentric
e. concentric

19. Which of the following substances mutates DNA by intercalatingbetween adjacent base pairs?

a. methyl methane sulfonate
b. all of these
c. nitrous acid
d. 2-amino urine
e. acridine orange

20. A molecule of RNA is always

a. double stranded
b. helical
c. nonparallel
d. all of these
e. none of these

21. Mutagenesis of cells using radiation or a chemicalintercalating agent could be used to create __________ mutations ina region of DNA.

a. random
b. both random and site-specific
c. site-specific

22. In Griffithsâs tranformation experiments, the mice died whenthey were injected with

a. living, no virulent bacteria
b. living, virulent bacteria
c. heat-killed, no virulent bacteria
d. living, nonvirulent bacteria
e. both B and D

23. A monoploid human cells

a. shows only the excessive phenotype of all these genes
b. would be functionally impaired
c. is normal
d. would not survive

24. Gametes containing an abnormal number of chromosomes may occuras a result of

a. paracentric inversion
b. reciprocal translocation
c. disjunction of chromosomes
d. unequal crossing over

25. DNA may be altered as a result of _____________ events thatoccur normally in cells

a. depurination
b. deamination
c. depyrimidation
d. all of the above
e. none of the above

26. Which enzyme prevents DNA from tangling up as the replicationfork migrates during replication?

a. DNA polymerase I
b. DNA helicase
c. topoisonmerase
d. DNA polymerase III
e. DNA gyrase

27. Which of the following enzymes play an important role in DNAreplication?

a. DNA helicase
b. DNA ligase
c. DNA primase
d. all of them
e. DNA polymerase III

28. A mutation gene sustains a mutation that restores the originalphenotype, This is a

a. frame shift mutation
b. transversion
c. reversion
d. transition
e. forward mutation

29. A dicentric chromosome has two

a. telomeres
b. mutations
c. arms
d. centromeres

30. Which type of chromosomal mutation cannot revert to thewild-type state

a. duplication
b. deletion
c. translocation
d. both A and C
e. all of them

31. Translation of histone mRNAs occurs during which phase of theeukaryotic cell cycle?

a. M
b. G1
c. G2
d. S
e. all of these

32. After removal of the RNA primers, Okazaki fragments are joinedthrough a reaction that involves which enzyme?

a. RNA polymerase
b. DNA helicase
c. DNA polymerase
d. DNA primase
e. DNA ligase

33. Which of the following is required for the synthesis of DNAduring replication?

a. DNA polymerase
b. magnesium ions
c. all of them
d. template DNA
e. dNTPs

34. Most of the genome of an active cell consists of

a. heterochromatin
b. histochromatin
c. poluchromatin
d. euchromatin

35. In eukaryotes, DNA replication begins,

a. a single origin of replication on each chromosome
b. a single origin of replication on each chromosome
c. multiple origins of replication on each chromosome
d. all of the above
e. none of the above

36. A human cell contains 2 sets of 24 chromosomes is

a. euploid
b. haploid
c. polyploid
d. aneuploid

37. Pseudodominance occurs as a result of which type ofmutation?

a. translocation
b. duppolication
c. all of these
d. point mutation
e. deletion

38. The enzymatic activity of a telomerase is best described asa

a. none of these
b. polymerase
c. ligase
d. reverse transcriptase
e. topoisomerase

39. A Barr body is an example of

a. constitutive heterochromatin
b. facultative heterochromatin
c. facultative euchromatin
d. constitutive euchromatin

40. An addition or deletion of one or two base pairs in a genecauses a ____________ mutation

a. Transition
b. frameshift
c. nonsense
d. transversion
e. neutral

41. A heterozygous chromosomal inversion can be detected

a. by cytogenetic observation of loops in chromosomes
b. through genetic studies indicating a decrease in the frequencyof recombination between genes
c. by observing a decrease in the number of viable progenyfollowing genetic crosses
d. all of the above

42. The mutagen 5-bromouracil is an analog of which base?

a. adenine
b. threonine
c. thymine
d. cystosine
e. uracil

43. A mutation that changes a codon from one that represents anamino acid to one that signals a chain termination is a______________ mutation.

a. neutral
b. missense
c. frame shift
d. nonsense

44. A chromosomal mutation that results in an increase in theamount of DNA is a(n)

a. duplication
b. inversion
c. deletion
d. both A and C
e. all of the above

45. Prior to replication, the DNA double helix must be untwisted bywhich type of enzyme?

a. topoisomerase
b. ligase
c. polymerase
d. helicase
e. primase

46. In his transformation experiments, Griffiths used cultures ofwhich bacterium?

a. Sacchromyces cerevisiae
b. Escherichia coli
c. Streptococcus pneumoniae.
d. Bacillus cereus
e. Eneterococcus faecalis

47. A mutation changes a codon from a UCG to a UAG. This is a____________ mutation?

a. point
b. nonsense
c. silent
d. both A and B
e. all of the above

48. In eukaryotes, DNA replication

a. is initiated at several sites on chromosomes
b. occurs bidirectional
c. is semidiscontinuous
d. all of the above
e. none of the above

49. Which of the following bacteria are used in the Amestest?

a. Pseudomonas aeruginosa
b. Escherichia coli
c. Vibrio cholera
d. Salmonella typhimurium
e. Rickettsia rickettsii

50. Which of the following changes represents a missensemutation?

a. UGA (stop) to UAG (stop)
b. AAA (lysine) to AGA (arginine)
c. UUU (phenylalanine) to UAA (stop)
d. AUG (methionine) to UGA (stop)
e. AAA (lysine) to AAG (lysine)

BIO 2306 Lecture Notes - Lecture 25: Frameshift Mutation, Sickle-Cell Disease, Replica Plating

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