Biotechnology Quiz: Restriction Enzymes & Cloning

The process of introducing foreign DNA into a host cell, causing the host to produce a desired protein or characteristic.

The process of using specific enzymes to cut DNA at precise locations, creating fragments for manipulation and analysis.

A specific DNA sequence recognized and cut by a restriction enzyme.

An enzyme that joins two DNA fragments together, forming a recombinant DNA molecule.

A self-replicating piece of DNA, often originating from bacteria or viruses, used as a vehicle to carry foreign DNA into a host cell.

A host cell that has been specially prepared to readily accept foreign DNA, often through treatment to increase membrane permeability.

A DNA molecule formed by combining DNA from different sources, often involving inserting foreign DNA into a vector.

A bacterial enzyme that removes phosphate groups from the 5' ends of DNA molecules, often used to prevent self-ligation of vectors during cloning.

A nomenclature system for restriction endonucleases, consisting of a 4-part code that defines the source bacterium, type of strain, and order of isolation.

Enzymes that cut DNA molecules at specific recognition sequences, creating either sticky or blunt ends.

REs produce DNA fragments with overhangs, often used for cloning and ligation.

REs produce DNA fragments with no overhangs, ideal for joining fragments with blunt ends.

Small circular DNA molecules found in bacteria that can replicate independently of the host chromosome. They are often used as vectors in genetic engineering to carry and replicate foreign DNA.

Viruses that infect bacteria and can be used as vectors in gene cloning. They are efficient at delivering their DNA into bacterial cells.

A type of cloning vector that combines features of plasmids and bacteriophages: contains a cos site (cohesive end sequence) that allows it to be packaged into phage particles.

The choice depends on the size of the DNA being cloned (gene of interest) and the type of host cell used ( prokaryotic or eukaryotic) for expression.

A type of cloning vector derived from bacteria, with a capacity of 350-500 kb for inserting foreign DNA.

A type of cloning vector derived from yeast, with a capacity of 1000 kb for inserting foreign DNA.

The origin of replication (ORI) controls how many copies of the plasmid are made, and it's where the foreign DNA is inserted. ROP is involved in repressing plasmid replication. Selectable markers, like antibiotic resistance genes, allow us to differentiate between transformed and non-transformed cells. Cloning sites, specific sequences for restriction enzymes, allow for the insertion of the foreign DNA.

Cells that have taken up the plasmid vector with the foreign DNA are called transformants. Non-transformants are the cells that didn't take up the plasmid.

Recombinant DNA contains the foreign DNA inserted into the vector, while non-recombinant DNA doesn't. We use techniques like antibiotic resistance screening to identify recombinants.

The Lac-Z gene codes for the enzyme β-galactosidase. When the foreign DNA is inserted into the Lac-Z gene, it disrupts the gene and the enzyme cannot be produced. This is called insertional inactivation. This technique allows for the identification of recombinant clones on a single plate.

Transformed cells with a non-recombinant plasmid can typically produce the β-galactosidase enzyme, which converts the X-gal substrate into a blue colored product. This results in blue colonies. On the other hand, transformed cells with a recombinant plasmid lack a functional β-galactosidase gene due to the insertion of the foreign DNA, resulting in white colonies.

pBR322 cloning vector was designed to test for antibiotic resistance against both ampicillin and tetracycline. When the foreign DNA is inserted into the tetracycline resistance gene (TetR), it disrupts this gene and the cell loses resistance to tetracycline. This allows for the identification of recombinants that are only resistant to ampicillin, but not tetracycline.

A specific DNA sequence recognized and cut by a restriction enzyme. The sequence is palindromic, meaning it reads the same backward as forward.

A specific DNA sequence recognized and cut by a restriction enzyme. The sequence is palindromic, meaning it reads the same backward as forward. Two strands of DNA have complementary restriction sites.

A technique used to separate DNA fragments based on their size and charge. DNA fragments are loaded into a gel and subjected to an electric field.

An enzyme that cuts DNA at specific sequences called restriction sites.

An enzyme that joins two DNA fragments together, forming a recombinant DNA molecule. It's like 'molecular glue' that seals DNA fragments together.

A self-replicating piece of DNA, often originating from bacteria or viruses, used as a vehicle to carry foreign DNA into a host cell for cloning.

A technique used to amplify specific DNA sequences exponentially.

A technique used to identify and quantify a specific protein in a sample by using antibodies.

The process of adding nucleotides to a growing DNA strand during DNA replication. This process is facilitated by enzymes like Taq polymerase at a temperature of 72°C, requiring magnesium ions (Mg++) for optimal activity.

A technique used to introduce foreign DNA into bacterial cells, where the cells are treated with a cold calcium chloride solution and then exposed to a brief heat shock.

In a recombinant DNA context, refers to the product of a gene that has been inserted into a different organism and is now being produced by that organism.

A large-scale vessel designed for cultivating microorganisms or cells in a controlled environment for the production of desired products.

A type of bioreactor with a simple design, often used in research and small-scale production. It features a stirred tank with features like a temperature sensor, pH monitor, foam breaker, and a flat blade impeller. This ensures efficient mixing and aeration of the cell culture.

A bioreactor equipped with a tube with many pores (spangen) to provide extra oxygen to the cell culture. This is crucial for efficient growth and production, especially when higher oxygen demands are present.

The final stage of recombinant DNA technology involves isolating the desired product (e.g., protein) from the cell culture, purifying it to remove unwanted impurities, and often adding preservatives to enhance stability. This prepares the product for commercial use.

A small molecule or segment of DNA used to identify a specific gene. It is labelled and used to detect complementary sequences in a sample.

A gene that allows identification of cells that have been successfully transformed (e.g., by incorporating a foreign gene). They usually confer resistance to a specific antibiotic.