Module 1.2: DNA Recombinant Technology

Question 1

Recombinant DNA are DNA molecules formed by laboratory methods of ____ (i.e., molecular cloning) in order to piece together genetic material from ____ sources

Answer 1

genetic recombination; multiple

Question 2

True or False. Recombinant DNA can be found naturally in the genome of organisms in nature.

Answer 2

False.

Question 3

Recombinant DNA technology comprises altering genetic material ____ an organism to obtain _____ characteristics in living organisms or as their products.

Answer 3

outside; enhanced and desired

Question 4

Recombinant DNA technology involves the insertion of ____ from a variety of sources, having a desirable gene sequence via the appropriate_____.

Answer 4

DNA fragments; vector

Question 5

Match the proponent/s for the discovery in the history of rRNA technology.

1. Was awarded the 1986 Nobel Prize for medicine in his discoveries on growth factors.
2. Developed the first recombinant DNA utilizing bacterial DNA and plasmids.
3. Conceived and approach to create rDNAs in vitro
4. Created the first chimeric DNA in vitro
5. Isolated the new cloning vector, pSC101 and created bacterial intra- and interspecies rDNAs

Choices:

Berg and colleagues
Berg and Loban
Herbert Boyer
Stanley Cohen
Jackson and colleagues

Answer 5

1. Stanley Cohen
2. Herbert Boyer and Stanley Cohen
3. Berg and Loban
4. Jackson and colleagues
5. Stanley Cohen and colleagues

Question 6

What was the first plasmid cloning vector?

Answer 6

lambda DVGal 120 (an operon encoding galactose metabolism genes concatenated with simian vacoulating virus 40)

Question 7

This hormone discovered by Stanley Cohen is involved in the proliferation and differentiation of cells in vivo.

Answer 7

Nerve growth factors and epidermal growth factors

Question 8

Memorize/familiarize

Answer 8

1. An approach to creating rDNA in vitro is conceived
2. The first bacterial cloning vector was isolated
3. First chimeric DNA is created
4. Second cloning vector is isolated
5. First intentional creation of rDNA molecules
6. Ribosomal genes propagated in E. coli cells
7. Asilomar conference looked at safety and containment issues of extrachromosomal DNAs and its risks of cancer
8. First gene transfer with humans
9. Gene therapy products are regulated by FDA
10. First approved gene transfer study on a chile with SCID
11. Gelsinger’s death
12. FDA elevates administrative unit that evaluates cellular, tissue, and gene therapy products.

Question 9

Genetic engineering refers to the process of using ____ to modify an organism’s ____ to achieve ____.

Answer 9

recombinant DNA technology; DNA; desirable traits

Question 10

Addition of ____ in the form of ____ that are generated by ____ is the most common method of genetic engineering

Answer 10

foreign DNA; recombinant DNA vectors; molecular cloning

Question 11

Other terms that are synonymous with genetic engineering

Answer 11

Recombinant DNA technology
Gene manipulation
Gene modification

Question 12

____ is the production of an exact copy—specifically, an exact genetic copy—of a gene, cell, or organism

Answer 12

DNA Cloning

Question 13

____ allows for the creation of multiple copies of genes, expression of genes, and study of specific genes

Answer 13

Molecular cloning

Question 14

_____ is a method used to make a clone or an identical copy of an entire multicellular organism.

Answer 14

Reproductive cloning

Question 15

____ is a combination of DNA fragments generated by ____ that does not exist in nature.

Answer 15

Recombinant DNA; molecular cloning

Question 16

Two types of enzymes in creating recombinant DNA.

Answer 16

restriction endonuclease; ligase

Question 17

Pipeline for recombinant DNA technology

Answer 17

1. Isolate the DNA sequence gene of interest
2. Insert DNA of interest into vector plasmid
3. Put recombinant plasmid into E. coli through the process of transformation
4. Clone cells
5. Choose colonies and inoculate a large culture to expand the plasmid carrying the gene of interest
6. Isolate and clean the DNA from the expanded culture
7. Transfer the plasmid DNA to desired cells and test the biological properties of the gene of interest

Question 18

During DNA isolation for the rDNA experiments, DNA is coexisting with other macromolecules within the cell membrane; hence, it must be separated and purified using enzymes such as

Answer 18

lysozymes, cellulase, chitinase, ribonuclease, and proteases

Question 18

During DNA isolation, DNA eventually precipitates out as fine threads as a result of the presence of ____.

Answer 18

ethanol

Question 19

During the cutting of DNA for an rDNA experiment, ____ helps to identify the location wherein a designated gene is introduced into a vector genome. The said reaction is known as ____. ____ is a technology that displays the restriction enzyme digestion’s progress

Answer 19

Restriction enzymes; restriction enzyme digestion; ‘Agarose Gel Electrophoresis’

Question 20

In the insertion to host phase, rDNA is added to the recipient host cell, and the entire process is called ____.

Answer 20

transformation (for bacteria)
transfection (for eukaryotes)
transduction (when a viral vector is used)

Question 21

foreign DNA. If so, they are given treatments to make them ‘capable’ of accepting new DNA. Give an example of this treatment.

Answer 21

Calcium ion treatments. Ca-DNA complex, endocytosis, proteolytic degradation of the endosome containing Ca-DNA complex

Question 22

In the recombinant cell isolation phase, only transformed host cells are filtered and isolated. The ___ of the plasmid vector is used to distinguish recombinant cells from non-recombinant cells.

Answer 22

marker gene

Question 23

Sequence the basic steps in recombinant DNA technology:

____ Recombinant Isolation
____ DNA isolation
____ Amplifying DNA
____ Insertion of rRNA into a host
____ Cutting of DNA/Restriction enzyme digestion
____ Joining DNA

Answer 23

6, 1, 3, 5, 2, 4

Question 24

Match the enzyme with its activity 1. plays a major role in determining the location at which the desired gene is inserted into the vector genome. 2. Cut within the DNA strand; scrutinize the length of DNA and make the cut at the specific site called the restriction site 3. Remove the nucleotides from the ends of the strands. 4. Synthesize DNA 5. Bind DNA sticky ends

Answer 24

1. Restriction enzymes 2. Endonuclease 3. Exonuclease 4. Polymerase 5. Ligases

Question 25

In rDNA technology, this refers to the component that helps in carrying and integrating the desired gene. It is also described as The ultimate vehicles that carry forward the desired gene into the host organism.

Answer 25

Vectors

Question 25

Match the type of polymerase to their activity 1. general PCR objectives 2. used for gene expression assay 3. Produces clean and high-yield amplicons 4. long PCR products

Answer 25

1. Taq polymerase 2. Proof-reading Taq polymerase 3. HotStart Taq 4. Long-range Taq NA polymerase

Question 26

Match the parts of a vector with its description. 1. a sequence of nucleotides from where the replication starts 2. constitute genes which show resistance to certain antibiotics like ampicillin; 3. sites recognized by the restriction enzymes wherein desired DNAs are inserted

Answer 26

1. origin of replication 2. selectable marker 3. cloning sites

Question 27

What are the two most common vectors for rDNA?

Answer 27

plasmid and bacteriophages

Question 28

This component of rDNA technology refers to where the recombinant DNA is introduced. It is the ultimate tool of recombinant DNA technology wherein it takes in the vector engineered with the desired DNA with the help of the enzymes.

Answer 28

Host organism

Question 29

Ways in which rDNAs are inserted into the host:

Answer 29

Microinjection Biolistics or gene gun Alternate cooling and heating Use of calcium ions Electrophoration

Question 30

Match the applications of recombinant DNA technology with its descriptions: 1. production of vaccines and protein therapies (i.e., human insulin, human growth hormone). 2. Used as an attempt to correct the gene defects which give rise to heredity diseases. 3. used as an attempt to correct the gene defects which give rise to heredity diseases. 4. Used to improve plant growth through insect and drought resistance and nutritional enrichment, by increasing nitrogen fixation efficiencies, cloning bacterial genes, and inserting them into plant cells.

Answer 30

1. DNA technology and pharmacies 2. Gene therapy 3. Clinical diagnosis 4. Agricultur

Question 31

Match the challenges and future prospects for rDNA technology. 1. More possible issues are cell stress responses activation, and instability of proteolytic activities, low solubility, and resistance in expressing new genes 2. Can cause deficiencies in proteins production 3. Possibility of genetically engineered plants cross-breeding with wild plants 4. Can make deadly pathogens (i.e., Y. pestis) resistant to modern antibiotics and can be fatal to humans

Answer 31

1. Posttranslational Modifications 2. Mutations Occurring In Humans At Genetic Levels 3. A Disaster For Safety And Biodiversity 4. Dangerous Health Implications

Question 32

The genetically modified tomato CGN-89564-2 was the first commercially grown, genetically engineered crop product to be granted a license for human consumption Developed in 1994 to express the trait of ____ of tomato flesh as a practical means to minimize post-harvest crop losses.

Answer 32

delayed softening

Question 33

____ an immunological assay commonly used to measure antibodies, antigens, proteins and glycoproteins in biological samples.

Answer 33

ELISA

Question 33

What are the strategies that have been used to treat cancers and diseases.

Answer 33

Inserting tumor suppressor genes oncolytic virotherapy gene directed enzyme prodrug therapy

Question 34

In the pharmacy industry, recombinant ____ was developed to support normal growth and development for patients with malfunctions in the ____.

Answer 34

human growth hormone; pituitary gland

Question 35

In the pharmacy industry, the ability to manufacture recombinant ____ allows larger quantities of blood; reduces the need for blood donation

Answer 35

blood clotting factor VIII

Question 36

In the pharmacy industry, recombinant DNA of the ___ is produced in yeast cells as part of the vaccine

Answer 36

hepatitis B virus surface antigen