Molecular Biology: Expression Vectors Quiz

True (A)

To control the expression of the cloned gene, allowing for the production of protein only when desired.

<p>expression vectors</p> Signup and view all the answers

<p>Integration into the host chromosome for stable inheritance (B)</p> Signup and view all the answers

<p>True (A)</p> Signup and view all the answers

<p>IPTG = Inducer molecule for T7 RNA polymerase pET vectors = Expression vectors based on the T7 expression system T7 RNA polymerase = A highly efficient RNA polymerase Fusion proteins = Proteins that combine two protein fragments for various purposes PolyHis tag = A peptide sequence used for protein purification using affinity chromatography lac promoter = Inducible promoter in bacteria that is activated by lactose or IPTG</p> Signup and view all the answers

<p>To efficiently transcribe cloned genes into mRNA for protein synthesis.</p> Signup and view all the answers

<p>post elution</p> Signup and view all the answers

<p>Resistance to all known antibiotics (C)</p> Signup and view all the answers

<p>False (B)</p> Signup and view all the answers

<p>The MCS contains a series of restriction enzyme recognition sites, allowing for the insertion of foreign DNA at a precise location within the plasmid.</p> Signup and view all the answers

<p>Small Size = Facilitates efficient entry into host cells and replication Selectable Markers = Allows for the selection of cells that have taken up the plasmid Unique Restriction Sites = Enable directional cloning and precise insertion of foreign DNA</p> Signup and view all the answers

<p>directional cloning</p> Signup and view all the answers

<p>True (A)</p> Signup and view all the answers

<p>Antibiotic resistance genes allow researchers to identify bacteria that have successfully taken up the plasmid containing the gene. Only bacteria with the plasmid will be able to grow in the presence of the specific antibiotic, thus selecting for transformed cells.</p> Signup and view all the answers

<p>All of the above (D)</p> Signup and view all the answers

<p>True (A)</p> Signup and view all the answers

<p>To transfer cloned DNA between unrelated organisms.</p> Signup and view all the answers

<p>His</p> Signup and view all the answers

<p>Shuttle vector = Can replicate in two different organisms. Expression vector = Designed for efficient protein expression. pMAL vector = Uses MBP fusion protein for purification.</p> Signup and view all the answers

<p>They always result in cytoplasmic expression of the protein (B)</p> Signup and view all the answers

<p>False (B)</p> Signup and view all the answers

<p>It guides the ribosome to the start codon for efficient translation.</p> Signup and view all the answers

<p>Chloramphenicol (B)</p> Signup and view all the answers

<p>True (A)</p> Signup and view all the answers

<p>The f1 origin allows for single-stranded DNA production.</p> Signup and view all the answers

<p>multiple cloning site (MCS)</p> Signup and view all the answers

<p>pUC origin = High copy number replication CAM resistance gene = Selection of transformed bacteria T7/T3 promoters = Production of strand-specific probes lacZ' gene = Blue/white screening for inserts</p> Signup and view all the answers

<p>All of the above (D)</p> Signup and view all the answers

<p>True (A)</p> Signup and view all the answers

<p>The blue/white screening technique helps identify bacterial colonies that contain the vector with an insert.</p> Signup and view all the answers

<p>multiple</p> Signup and view all the answers

<p>Ampicillin resistance gene (A)</p> Signup and view all the answers

<p>To identify bacteria containing recombinant plasmids (C)</p> Signup and view all the answers

<p>True (A)</p> Signup and view all the answers

<p>ampR and lacZ</p> Signup and view all the answers

<p>transformation</p> Signup and view all the answers

<p>pGEM series = Promega pGEX series = Pharmacia pKK223-3 = Pharmacia pMAL = New England Biolabs</p> Signup and view all the answers

<p>Insertion of DNA into the vector (C)</p> Signup and view all the answers

<p>False (B)</p> Signup and view all the answers

<p>Nucleic acid hybridization</p> Signup and view all the answers

<p>To bind to DNA (C)</p> Signup and view all the answers

<p>False (B)</p> Signup and view all the answers

<p>12.0-12.5</p> Signup and view all the answers

<p>Tris·Cl–EDTA</p> Signup and view all the answers

<ol> <li>Lyse cells = A. Neutralizes the lysate and causes precipitation of contaminants</li> <li>Neutralize lysate = B. Releases the cell contents and denatures DNA</li> <li>Clear the lysate = C. Removes insoluble debris through centrifugation or filtration</li> <li>Resuspend cells = D. Prepares samples for lysis in appropriate buffer</li> </ol> Signup and view all the answers

<p>Silica supports binding in the presence of chaotropic salts (A)</p> Signup and view all the answers

<p>False (B)</p> Signup and view all the answers

<p>To neutralize the lysate</p> Signup and view all the answers

Covalently closed circular DNA, like plasmid DNA, has no free ends.

DNA with free ends, such as fragmented chromosomal DNA, allows more binding of EtBr.

A dye that binds to DNA; more binding occurs in linear DNA than CCC DNA.

Popular technique for isolating plasmid DNA, using NaOH at pH 12.0-12.5.

Resuspend bacterial cells in Tris·Cl-EDTA buffer containing RNase A.

Lyse cells using NaOH/SDS to release cell contents and denature DNA.

Conditions allowing DNA to bind include low salt or chaotropic salts.

Separating plasmids from solid support using high salt buffer or water.

A method for identifying recombinant bacteria using color change in the presence of X-gal.

A process involving five main steps to clone a gene into a plasmid.

The first step in cloning where the vector and gene-source DNA are collected.

An enzyme that cuts DNA at specific recognition sequences.

The process by which bacteria take up recombinant plasmids.

A technique to differentiate between recombinant and non-recombinant bacteria based on colony color.

A method to identify desired genes using complementary probes.

DNA formed by combining genes from different sources.

The process of separating the DNA strands in a plasmid.

Plasmid vectors can carry up to 10 kb of inserted DNA only.

Genetically modified lambda phage used for larger DNA fragments.

Essential features for plasmid vectors to express genes effectively.

Plasmids that can exist in many copies to enhance gene dosage.

Expression is regulated with specific promoter constructs to optimize growth.

Proteins created from joining two or more genes to increase stability and ease of detection.

The phase where expression of the gene is induced after cell growth.

Process of removing salt from plasmid using isopropanol.

A cloning vector should be small, selectable, and have unique sites.

Genes that allow identification of transformed cells, usually antibiotic resistance.

Common markers include genes for resistance to ampicillin, tetracycline, chloramphenicol, and kanamycin.

A widely used plasmid cloning vector in E. coli with a medium copy number.

A region with consensus sequences for various restriction enzymes for DNA insertion.

Inserting DNA in a specific orientation using two different restriction enzymes.

The process of identifying bacteria with the recombinant plasmid containing the DNA of interest.

Experimental methods used to study binding interactions, such as protein-ligand or protein-protein binding.

A process used to identify and select specific antibodies from a pool, which can bind to particular antigens.

Sequences in proteins that direct their transport to specific cellular locations, such as the periplasm or outside the cell.

A protein used in affinity purification linked to maltose, allowing recovery of fusion proteins.

A sequence of six histidine residues added to proteins, allowing them to bind metal for purification.

Plasmids designed to increase expression of cloned genes in host organisms, aiding protein production.

Vectors that can replicate in two different organisms, allowing gene transfer between species.

A strong hybrid promoter used in certain vectors to drive high levels of gene expression.

Origin of replication that allows high copy number.

Gene that provides resistance to the antibiotic chloramphenicol.

A short segment of DNA containing multiple restriction sites.

A method to identify recombinant bacteria based on color.

Single-stranded DNA production through f1 origin.

Sequences that initiate transcription for making probes.

A gene segment used for blue/white screening in cloning.