🏷️ LIMITED TIME OFFER: GET 20% OFF GRADE+ YEARLY SUBSCRIPTION →
Log in
HomeClass Notes1,200,000CA670,000

MBB 222 Lecture Notes - Lecture 23: Microsatellite, Acrylamide, Chromosome

43 views4 pages
blacktortoise173
21 Mar 2016
School
Simon Fraser University
Department
Molec Biol & Biochem
Course
MBB 222
Professor
dr.chu

For unlimited access to Class Notes, a Class+ subscription is required.

Document Summary

Gene encoding a protein that causes the cell to produce a easily identifiable, detectable product (usually coloured or fluorescent). e. g. blue/white screening involving lacz gene using the pbluescript plasmid. Can use expression vectors to express the gene product (protein). Can modify the proteins of interest for affinity purification procedure, or even visualization. Can introduce favourable genes into plants and animals (transgenics). Can make many copies of the gene of interest (pcr). Can determine the nucleotide sequence of the gene (sequencing). Can modify the gene at specific sites to give the protein different properties (site directed mutagenesis). Can create genomic or cdna libraries (dna libraries). Typical cloning vector features: bacterial origin of replication, selectable marker (antibiotic resistance, polylinker (multiple cloning site, screening markers. System for regulating transcription of inserted dna. Transcription termination sequence at end of insert. Can contain sequences to allow production of fusion or tagged proteins.

Get access

Grade+20% off
$8 USD/m$10 USD/m
Billed $96 USD annually
Grade+
Homework Help
Study Guides
Textbook Solutions
Class Notes
Textbook Notes
Booster Class
40 Verified Answers
Class+
$8 USD/m
Billed $96 USD annually
Class+
Homework Help
Study Guides
Textbook Solutions
Class Notes
Textbook Notes
Booster Class
30 Verified Answers

Related Documents

MCB 2050
Midterm Exam
Study Guide
MCB 2050 Study Guide - Comprehensive Midterm Guide: Apoptosis, Plant Cell, Thymidine
jadeant325
MCB 2050 Lecture Notes - Lecture 15: International Hapmap Project, Avidin, Embryonic Stem Cell
jadeant325
MCB 2050
Final Exam
Study Guide
MCB 2050 Study Guide - Midterm Guide: Polymerase Chain Reaction, Multiple Cloning Site, Agarose Gel Electrophoresis
olivegoat3

Related Questions

Please answer the following multiple-part question regarding vectored cloning:

a) A cloning vector must _________.

A. be a circular dsDNA molecule

B. contain a replication origin

C. contain an multicloning site (MCS)

D. contain marker genes

b) Vectored cloning is superior to PCR cloning of a gene in terms of __________.

A. time it takes to finish cloning

B. size of DNA that can be cloned

C. amounts of cloned DNA that can be generated

D. ease of isolation of the DNA fragment to be cloned

c) Which of the following is a recombinant DNA molecule?

A. pUC DNA digested by EcoRI

B. Fish insulin gene, PCR amplified and then cut by EcoRI

C. pUC18 DNA and fish insulin DNA ligated by a DNA liagse

D. pUC DNA and fish insulin gene DNA, both cut by EcoRI and mixed

d) You may insert a foreign DNA fragment inside a live cell using each of the following methods except________.

A. inject the DNA inside the cell

B. mix the DNA with the cell and freeze the mixture in liquid nitrogen

C. mix DNA with the cell and momentarily heat the cell then cool down

D. mix DNA with the cell and momentarily zap the cells with a high voltage

E. cover the DNA in a lipid micelle and mix the micelle with the cell

e) Technically, the ampicillin resistance gene is a selectable marker gene but the beta lacZ gene or the green fluorescent protein (GFP) gene is a reporter gene of a cloning vector. What is the difference between a selectable marker and a reporter gene?

A. A selectable marker is a bacterial gene, a reporter gene is a eukaryotic gene

B. Marker genes are used in bacterial system; reporter genes are used in eukaryotic system.

C. The selectable markers help eliminate cells lacking the maker, reporter genes do not offer this option.

Which of the following did Erwin Chargaff observe?

A)

Base composition changes as the organism ages.

B)

Base composition of DNA does not vary from one species toanother.

C)

In all species the number of adenines equals the number ofguanines.

D)

In all species A + T = G + C.

E)

DNAs isolated from different tissues of the same species havethe same base composition.

The melting temperature (Tm) of a DNA duplex is:

A)

the temperature at which double-stranded nucleic acids degradeinto free nucleotides.

B)

the temperature at which double-stranded DNA is toosingle-stranded to ever renature back to double strands.

C)

the temperature at which double-stranded DNA is completelymelted.

D)

the temperature at which double-stranded DNA has become 50%single-stranded.

E)

the temperature at which double-stranded DNA begins to melt.

Given your knowledge and the descriptions of each of thesemethods, which of the following does NOT rely on the ability ofnucleic acids to hybridize with each other?

A)

Southern blotting for the detection of specific DNA fragmentsthat have been separated by gel electrophoresis.

B)

Colony blot hybridization to identify a specific sequence from acollection of sequences that have been inserted into bacteria.

C)

Northern blotting for the detection of specific RNA fragmentsthat have been separated by gel electrophoresis.

D)

A DNA oligonucleotide microarray incubated with probes made fromcDNA.

E)

Western blotting for the detection of specific proteins thathave been separated by gel electrophoresis.

Which of the following is a possible reason why DNA uses thymineinstead of uracil?

A)

Cytidine is deaminated to uracil so it would be difficult forany repair mechanism to differentiate between uracil that belongsin the sequence and uracil that resulted from deamination.

B)

Adenine is deaminated to uracil so it would be difficult for anyrepair mechanism to differentiate between uracil that belongs inthe sequence and uracil that resulted from deamination.

C)

When uracil is bound to a deoxyribose it will become morereactive, forming covalent bonds with other bases.

D)

None of the above.

Which of the following is an enzyme used in cloning to breakcovalent bonds?

A)

DNA ligase

B)

Restriction endonuclease

C)

Reverse transcriptase

D)

DNA polymerase I

E)

Alkaline phosphatase

Which of the following could you use to remove the 5' phosphatesafter cleavage of a plasmid with a single type of restrictionenzyme to ensure that the plasmid would not simply fuse backtogether upon addition of ligase?

A)

Reverse transcriptase

B)

Alkaline phosphatase

C)

DNA ligase

D)

DNA polymerase I

E)

Restriction endonuclease

Which of the following would you use to create a cDNA librarythat you would not use for a genomic library?

A)

Alkaline phosphatase

B)

Reverse transcriptase

C)

Restriction endonuclease

D)

DNA ligase

E)

DNA polymerase I

A plasmid vector typically has which of the followingfeatures?

A)

An origin of replication

B)

A selectable marker to ensure that the only bacteria growingcontain the intact or recombinant vector.

C)

Unique restriction sites for insertion of DNA.

D)

Small size to facilitate entry into the cell and biochemicalmanipulation of the DNA.

E)

All of the above are features of plasmid vectors.

Which of the following is FALSE of genomic or cDNAlibraries?

A)

Genes represented in cDNA libraries include control sequencesthat direct transcription.

B)

cDNA libraries contain cDNAs made from mRNA.

C)

Genomic libraries contain fragments of genomic DNA that aregenerated by partial digests with restriction enzymes.

D)

Genomic libraries usually contain large fragments and are likelyto be constructed in YAC or BAC vectors.

E)

cDNA libraries contain sequences from expressed genes only.

Which of the following is NOT a typical component of apolymerase chain reaction (PCR)?

A)

dNTPs

B)

DNA containing the sequences to be amplified

C)

DNA ligase to connect the fragments together

D)

Primers complementary to each end of the sequence to beamplified

E)

Heat stable Taq polymerase

How can a fusion protein be formed?

A)

Site-directed mutagenesis where a fragment is replaced with asynthetic sequence containing the altered DNA sequence.

B)

Two proteins can recombine in the bacterial cell to form a newhybrid protein.

C)

Portions of two different genes are ligated together to create anew hybrid gene. When expressed the gene product is a fusion of thetwo gene products.

D)

Using oligonucleotide-directed mutagenesis to introduce specificmutations that cause the expressed protein to fuse to otherproteins.

E)

A portion of the gene can be removed to form a shorter gene andtherefore a shorter protein.

Why is green fluorescent protein (GFP) so useful in visualizingfusion proteins in eukaryotes?

A)

The GFP protein recognizes and binds to fusion proteins allowingthe location of the fusion protein to be determined.

B)

The reaction is only dependent on the presence of GFP and oneother cofactor.

C)

It is easily cloned and expressed in bacterial cells.

D)

The presence of just a few molecules of GFP fusion proteins canbe sufficient for observing the proteins microscopically allowingthe study of its location and movements in the cell.

E)

It is derived from fire flies, which are easy to cultivate inthe lab.

You wish to determine the cellular location of a protein butunfortunately it is not particularly antigenic (can t develop astrong antibody response). How could you solve this problem?

A)

Create a fusion protein with an epitope tag that can bevisualized by incubation with fluorescently tagged antibody to theepitope.

B)

Create a fusion with green fluorescent protein (GFP).

C)

Express the protein as a fusion with biotin. The addition of GFPwill cause fluorescence.

D)

A and B might work.

E)

A, B, and C might work.

F)

None of the above will work.

A positive result for the yeast two-hybrid analysis means thefollowing:

A)

The proteins that you are studying can be made into fusionproteins, but do NOT interact with each other within the nucleus ofyeast.

B)

A fusion protein containing the Gal4p activation domain, hasbound to RNA polymerase resulting increase transcription ofnumerous genes, including the reporter gene, and formation ofcolonies.

C)

Two fusion proteins, one containing the Gal4p binding sitedomain and one containing the Gal4p activation domain, haveinteracted through their Gal4p domains resulting in transcriptionof the reporter gene.

D)

Two fusion proteins interact with each other and because of theGal4p binding site domain and the Gal4p activation domain containedin each fusion protein, the reporter genes are activated and thecells die resulting in no colonies.

E)

The colonies growing are expressing the two fusion proteins, butthe two proteins have not interacted with each other.

Which of the following is the approximate size of the humangenome?

A)

3 Gm

B)

300 Kb

C)

3 Mb

D)

3 Gb

E)

3 Mm

Which of the following is correct about the structure orlocation of genes?

Question 22 options:

A)

Prokaryotic genes typically possess their own regulatorysequences that only control expression of a single gene.

B)

Eukaryotic genes often encode introns that are spliced outbefore translation.

C)

Eukaryotic genes are mostly located at the telomeres and thecentromeres of chromosomes.

D)

Eukaryotic genes are typically arranged in operons.

E)

Prokaryotic genes often encode exons that are spliced out beforetranslation