p.8
Amplification of Gene of Interest using Polymerase Chain Reaction (PCR)
What laboratory method is used to make many copies of a specific piece of DNA?
Polymerase Chain Reaction (PCR).
p.12
Tools and Techniques of Genetic Engineering
Which enzymes are used in recombinant DNA technology or genetic engineering?
Restriction endonucleases and DNA ligase.
p.2
Recombinant DNA Technology
Who introduced the gene of the SV 40 virus into E.coli?
Paul Bergh (Father of Genetic Engineering) with the help of the phage.
p.4
Tools and Techniques of Genetic Engineering
What are the enzymes used to make new DNA strands complementary to a DNA or RNA template?
Reverse transcriptases and DNA polymerases.
p.2
Tools and Techniques of Genetic Engineering
What is the function of exonucleases in genetic engineering?
To cut off nucleotides from 5' or 3' ends of DNA molecules.
p.4
Tools and Techniques of Genetic Engineering
What do joining enzymes aid in?
The recombination of DNA fragments.
p.3
Recombinant DNA Technology
How is a recombinant DNA molecule created using restriction enzymes?
By cutting two distinct DNA samples with the same restriction enzyme and combining the fragments together.
p.9
Process of Recombinant DNA Technology
What happens during the denaturation phase of the PCR cycle?
Double-stranded DNA is heated to 94°C to separate the two strands.
What is the significance of the EcoRI restriction enzyme?
It was identified by Arber, Smith, and Nathans and is utilized in recombinant DNA technology.
p.7
Process of Recombinant DNA Technology
What are the three goals for creating recombinant DNA molecules?
To get a lot of copies of a certain DNA segment, to recover a large amount of the protein generated by the gene, and to integrate the gene into a target organism's genome.
p.9
Process of Recombinant DNA Technology
What occurs during the annealing phase of the PCR cycle?
Primer binding to the 3' end of single strands of DNA at a decreased temperature of 56°C.
p.2
Tools and Techniques of Genetic Engineering
What is the function of lysing enzymes in genetic engineering?
To open cells in order to obtain DNA for genetic experiments.
p.9
Process of Recombinant DNA Technology
What are the three steps of the PCR cycle?
1. Denaturation, 2. Annealing, 3. Extension.
Which enzyme is used to connect DNA fragments?
DNA ligase from Escherichia coli.
How does genetic engineering alter DNA?
Artificially and purposefully, to meet human requirements.
p.7
Process of Recombinant DNA Technology
What is the purpose of genetic engineering in the process of creating recombinant DNA molecules?
To isolate the desired gene from any creature and then transfer and express it into the organism of choice.
p.7
Process of Recombinant DNA Technology
What is an example of a therapeutic protein produced from transgenic microbes?
Human insulin, commercially produced from a transgenic E.coli strain.
Why are antibiotic resistance genes considered valuable selection markers for E. coli?
They allow for the selection of transformed cells by growing them on media containing specific antibiotics.
p.6
Tools and Techniques of Genetic Engineering
What is random DNA?
Tiny pieces created when restriction endonucleases are used to break a chromosome.
p.8
Isolation of Specific Genetic Material (DNA)
What enzymes are used to treat bacterial cells, plant cells, or animal tissue during the isolation process?
Enzymes such as lysozyme, cellulase, and chitinase.
p.4
Tools and Techniques of Genetic Engineering
What is the function of DNA polymerases?
To help in the synthesis of complementary DNA strands on DNA templates.
p.1
Traditional Biotechnology
Give an example of a product produced by traditional biotechnology.
Citric acid by Penicillium notatum, and Penicillin by Penicillium notatum.
What are plasmids?
Extra chromosomal DNA segments found in bacteria that may replicate independently.
p.9
Process of Recombinant DNA Technology
What happens during the extension step of the PCR cycle?
Enzyme Taq Polymerase extends the primers by adding nucleotides to the 3' end until a complementary DNA strand is formed.
p.11
Recombinant DNA Technology
What does the PCR cycle involve?
Three steps: Denaturation, Annealing, and Extension.
What is the purpose of cloning sites in a vector?
To provide recognition sites for restriction enzymes to link the alien DNA.
p.5
Recombinant DNA Technology
What is the role of T-DNA in Agrobacterium tumefaciens?
It is used to turn normal plant cells into tumours.
p.4
Tools and Techniques of Genetic Engineering
What is the function of reverse transcriptases?
To help in the synthesis of complementary DNA strands on RNA templates.
p.1
Traditional Biotechnology
What did traditional biotechnology rely on?
Microorganism's inherent abilities.
What is the pBR vector plasmid named after?
The discoverer Bolivar and Rodriguez.
p.6
Process of Recombinant DNA Technology
How were the genes for galactosidase transferred from Escherichia coli to human cells?
Using a bacteriophage (bacterial virus).
How are recombinants distinguished from non-recombinants in the presence of two antibiotics?
Recombinants grow on one antibiotic but not the other, while non-recombinants grow on both antibiotics.
p.1
Biotechnology Definition
How is biotechnology defined?
As the use of microorganisms, animals, or plant cells, or their products, to produce valuable products and services on an industrial scale.
p.3
Tools and Techniques of Genetic Engineering
What is the most widely used medium in laboratories for bacteria culture and gel electrophoresis?
Agarose, a natural polymer produced from seaweeds.
p.8
Cutting of DNA at specific locations
What is the function of DNA ligase?
To bind two pieces of DNA together, creating recombinant DNA.
p.1
Recombinant DNA Technology
What is the heart of new biotechnology?
Recombinant DNA technology.
p.11
Traditional Biotechnology
What are the two ways microorganisms can be grown in bioreactors?
Support growth system and Suspended growth system.
p.7
Application of Recombinant DNA Technology
Name some applications of recombinant DNA technology?
Decipher molecular events in biological processes, create useful chemical compounds, produce transgenic plants and microorganisms.
What is the purpose of using a chromogenic substrate in selecting recombinants?
To distinguish recombinant colonies from non-recombinant colonies based on their ability to produce color.
p.8
Recombinant DNA Technology
What process involves the removal of plasmids or genomic DNA from cells?
Isolation of Specific Genetic Material (DNA).
p.12
Tools and Techniques of Genetic Engineering
What happens in the extension step of PCR?
The temperature of the reaction is raised to 72°C and the primers are extended by the enzyme Taq Polymerase.
p.10
Recombinant DNA Technology
What is the purpose of expressing foreign genes in Recombinant DNA technology?
To obtain the foreign gene product.
p.10
Application of Recombinant DNA Technology
What kind of conditions do bioreactors provide for producing the desired product?
Optimal growth conditions such as temperature, pH, substrate, salts, vitamins, and oxygen.
What is meant by 'Ori' in the context of cloning vector?
Replication's starting point - the sequence in which replication begins.
What is the downstream process in biotechnology?
A series of processes including separation, purification, and addition of preservatives after the biosynthetic stage.
p.5
Tools and Techniques of Genetic Engineering
What is a selectable marker in genetic engineering?
It is a gene that aids in the selection of successfully transformed cells.
p.11
Recombinant DNA Technology
What happens during the Annealing phase of the PCR cycle?
Primers, 20 base pairs (bp) long, bind to the 3' end of single strands of DNA.
p.5
Recombinant DNA Technology
How do retroviruses impact cells in animals?
They have the power to change healthy cells into malignant ones.
p.10
Recombinant DNA Technology
What is the ultimate goal of Recombinant DNA technology?
To create a desirable protein.
What is the function of restriction endonucleases in genetic engineering?
To cleave DNA duplexes at specific points in such a way that they come to possess short single-stranded free ends.
p.7
Process of Recombinant DNA Technology
What is a recombinant DNA molecule?
A molecule created when two or more DNA segments from different organisms are joined together.
What is the function of alkaline phosphatases in cloning?
To prevent recircularization by removing a phosphate group from the 5' end of linearized circular DNA.
p.10
Tools and Techniques of Genetic Engineering
How do stirred-tank reactors aid in the mixing of reactor contents?
They ensure even mixing and availability of oxygen at all times.
p.11
Traditional Biotechnology
What is the purpose of a very large-sized bioreactor in manufacturing?
To accommodate a huge amount of medium for the process.
p.11
Recombinant DNA Technology
Describe the Denaturation phase of the PCR cycle.
Double-stranded DNA is heated to 94°C for two minutes to separate the two strands.
What is insertional inactivation?
It is the loss of antibiotic resistance in recombinant plasmids due to the inclusion of foreign DNA.
What are the characteristics of DNA fragments produced by restriction enzymes?
They contain brief single-stranded overhangs at each end, referred to as sticky or cohesive ends.
p.8
Cutting of DNA at specific locations
How is DNA cleavage made feasible?
By the use of restriction endonucleases.
p.12
Recombinant DNA Technology
What is the modern biotechnology process that uses genetically modified organisms to alter the DNA's chemistry?
Recombinant DNA technology or genetic engineering.
p.10
Application of Recombinant DNA Technology
What are bioreactors used for in genetic engineering?
To process enormous volumes of culture and manufacture products in large quantities.
p.6
Tools and Techniques of Genetic Engineering
How can plasmids be extracted from bacteria and joined with appropriate DNA segments?
Using restriction enzymes and DNA ligase.
p.7
Process of Recombinant DNA Technology
What are transgenic organisms?
Organisms where the desired gene is transferred and expressed, known as transgenic organisms.
p.7
Process of Recombinant DNA Technology
What are transgenic animal cell lines and transgenic plants used for?
To make a variety of useful recombinant proteins.
What is passenger DNA?
The DNA combined with the vehicle DNA and transported from one organism to another.
p.5
Recombinant DNA Technology
What has been transformed into useful vectors for delivering genes of interest to humans?
Pathogens' tools for delivering genes to their eukaryotic hosts.
What is used to isolate foreign DNA into host organisms in recombinant DNA technology?
Plasmid or viral vectors.
p.1
Tools and Techniques of Genetic Engineering
What technique is used in genetic engineering to manipulate DNA?
Breaking a DNA molecule at two specified locations with restriction endonuclease to extract a specific DNA segment and then inserting it into another DNA molecule at a desired position.
What is the goal of genetic engineering?
To add, remove, or repair a piece of genetic material.
p.9
Recombinant DNA Technology
What is the commonly used host for the introduction of foreign DNA?
Escherichia coli (E. coli).
How is foreign DNA ligated with the vector using a restriction site?
A restriction site located in one of the antibiotic resistance genes is used to ligate foreign DNA.
What are recombinant plasmids, hybrid plasmids, and chimeric plasmids?
Plasmids that have the DNA of another organism integrated into them.
p.7
Process of Recombinant DNA Technology
What does the recombinant DNA technique entail?
A series of steps that must be completed in a certain order, such as isolation of a particular type of genetic material.
p.7
Process of Recombinant DNA Technology
What are recombinant proteins?
Proteins created by transgenes and used to create a variety of products.
p.6
Tools and Techniques of Genetic Engineering
How is complementary DNA (cDNA) created?
Using reverse transcriptase and the required nucleotides on an mRNA template.
