Dela Peña, Chinee T.

03/14/22
11 - Ilmenite

General Biology 2
Activity 1: Genetics
I. Define the following terms:

1. Genetic Engineering - Genetic engineering, a.k.a. genetic modification,

is the process of manipulating the DNA in an organism's genome. Genetic

engineering is used by scientists to enhance or modify the characteristics

of an individual organism.

2. DNA - DNA stands for deoxyrubonucleic acid; it is the long molecule that

contains the unique genetic code of all living things.

3. Recombinant DNA - is the method of joining two or more DNA molecules

to create a hybrid. The technology is made possible by two types of

enzymes, restriction endonucleases and ligase.

4. Plasmids - These small circles of genetic material,

typically DNA in bacterial cells, physically separated from

chromosomal DNA that replicates independently.

Plasmids are used as vectors to transfer genes between

cells.

5. Cloning - It is the natural or artificial process of

creating an exact copy (clone) of whole organisms or

cells but also molecules such as DNA.

6. Genome - A genome is an organism’s complete set of genetic

instructions. Each genome contains all of the information

needed to build that organism and allow it to grow and develop.

7. Gene Mapping - Genome mapping is used to identify and

record the location of genes and the distances between genes

on a chromosome.
 8. Biotechnology - Biotechnology is the use of living

organisms and biological processes to make

commercially useful products or services.

9. Polymerase Chain Reaction - First developed in the 1980s, it is a technique

that is used to in molecular biology to make many copies of small sections of

DNA or a gene. It is used in the early stages of processing DNA for

sequencing, for detecting the presence or absence of a gene to help identify

pathogens during infection, and when generating forensic DNA profiles from

tiny samples of DNA.

10. Gene Therapy - Gene therapy is when DNA is

introduced into a patient to treat a genetic disease.

The new DNA usually contains a functioning gene to

correct the effects of a disease-causing mutation.

 References

https://www.yourgenome.org/

https://www.sciencedirect.com/

https://www.genome.gov/
Dela Peña, Chinee T.
II. Answer the following:
11 - Ilmenite

1. How can organisms be modified?

Organisms may be genetically modified through different methods

and techniques.

Genetic Engineering Techniques

In Plants In Animals
Recombinant DNA Technology Recombinant DNA Technology

Cloning
Cloning
Accessing the Germline of Animals
Microbial Vectors
Transfection
Microprojectile Bombardment
Retroviral Vectors
Electroporation
Transposons( not completely developed yet)

Microinjection Knock-In and Knock-Out Technology

Transposons/Transposable Elements Marker-Assisted Selection

Nontransgenic Methods of Animal

Nontransgenic Molecular Methods of
Manipulation
Manipulation
Dela Peña, Chinee T.

11 - Ilmenite

Modifying Techniques Other than Genetic Engineering

In Plants In Animals
Domestication and Artificial Selection
Simple Selection
Assisted Reproductive Procedures
Crossing

Interspecies Crossing
Genetic Modification of Microbes
Embryo Rescue
Traditional genetic modification methods that have
Somatic Hybridization
been employed—particularly for microbial starter
Somaclonal Variation
cultures—include selection, mutagenesis,
Mutation Breeding: Induced
conjugation, and protoplast fusion. Before

Chemical and X-ray Mutagenesis molecular genetics was developed and applied to

Cell Selection LAB, the most widely used genetic modification

method was chemical- or ultra-violet-induced

Reference:
mutagenesis, followed by an enrichment or selection
https://www.ncbi.nlm.nih.
process for mutants with superior characteristics.
gov/books/NBK215771/
2. Enumerate 5 plants or animals that have desirable or enhanced traits and

how each of the traits was introduced or developed.

ENHANCED TRAIT: Bt Corn (insect pests resistant, specifically the European corn borer)
1.
MODIFYING TECHNIQUE: Recombinant DNA Technology
Bacillus thuringiensis, commonly known as Bt, is

a bacterium that occurs naturally in the soil.

For years, bacteriologists have known that

some strains of Bt produce proteins that kill

certain insects with alkaline digestive tracts.

When these insects ingest the protein

produced by Bt, the function of their digestive

systems is disrupted, producing slow growth

and, ultimately, death.

In 1983, the World Health Organization used Bt in West Africa to control disease-

carrying blackflies. In the U.S., various strains of Bt are used to control spruce

budworms and gypsy moths in forests, cabbage worms in broccoli and cauliflower,

loopers or budworms in cotton and tobacco, and leaf rollers in fruits.

However, less than one percent of all pesticides used in the U.S. each year contain Bt

(Monsanto). As an ingredient of commercial sprays, Bt is relatively expensive and has

some drawbacks. Although some pesticides kill on contact, Bt must be eaten by

insects to be effective. Sunlight breaks down Bt, and rain washes it from the plants.

Therefore, Bt must be applied exactly where and when the target insects are feeding,

and they must consume it quickly before it disappears.

 Figure 1
In the last twenty years, scientists made a

surprising discovery — DNA is interchangeable

among animals, plants, bacteria ... any

organism! In addition to using traditional

breeding methods of improving plants and

animals through years of cross-breeding and

selection, scientists can now isolate the gene

or genes for the traits they want in one animal

or plant and move them into another.

DNA technology makes it possible to locate

the gene that produces Bt proteins lethal to

insects and transfer the gene into crop plants.

The process is depicted in Figure 1.

First, scientists identify a strain of Bt that kills the targeted insect. Then they isolate the gene that

produces the lethal protein. That gene is removed from the Bt bacterium, and a gene conferring

resistance to a chemical (usually antibiotic or herbicide) is attached that will prove useful in a

later step.

The Bt gene with the resistance gene attached is inserted into plant cells. At this point, scientists

must determine which plant cells have successfully received the Bt gene and are now

transformed. Any plant cell that has the Bt gene must also have the resistance gene that was

attached to it. Researchers grow the plant cells in the presence of the antibiotic or herbicide

and select the plant cells that are unaffected by it. These genetically transformed plant cells are

then grown into whole plants by a process called tissue culture. The modified plants produce the

same lethal Bt protein produced by Bt bacteria because the plants now have the same gene.

Research to transfer insect resistance genes from Bt to crop plants is well under way. Corn,

cotton and potatoes are three of the many commercial crops targeted for Bt insect resistance.

Reference:https://extension.missouri.edu/publications/ncr553
2. ENHANCED TRAIT: Golden Rice (Vitamin Enrichment)
MODIFYING TECHNIQUE: Recombinant DNA Technology
Golden Rice was engineered

from normal rice by Ingo Potrykus

and Peter Beyer in the 1990s to

help improve human health.

Golden Rice has an engineered

multi-gene biochemical pathway

in its genome. This pathway

produces beta-carotene, a

molecule that becomes vitamin A

when metabolized by humans.

Picture retrieved from

Reference: https://www.isaaa.org/kc/inforesources/biote
https://embryo.asu.edu/pages/gol
chcrops/the_golden_rice_technology.htm
den-rice
Golden Rice is named for its golden color, which is caused by beta-carotene. Normal rice, Oryza sativa,

does not express beta-carotene in its endosperm—the starchy and biggest part of the rice seed, which is

usually an off-white color. Beta-carotene is part of a class of molecules called carotenoids, one of

hundreds that plants naturally produce, and it has a yellow-orange hue. Carotenoids are essential

nutrients for humans, because they are precursors to molecules needed in metabolism. The human body

transforms beta-carotene, also known as pro-vitamin A, into vitamin A, which is necessary to produce

retinal and retinoic acid. When people lack access to foods containing beta-carotene, because they eat

mostly cereal crops such as rice, wheat, or sorghum, they are at risk of blindness and disease.

Because no one had previously successfully expressed three genes in a food crop, Potrykus' lab

attempted multiple methods for the transformation. The first step was to insert the genes into the rice

embryo, through particle bombardment or bacterial transfer. Potrykus' lab used an Agrobacterium-

mediated transformation, where engineered bacteria inserted its DNA into the targeted rice plant

embryos. This DNA contained all three genes—phytoene synthase (psy, from daffodil), phytoene

desaturase (crtI from bacteria), and lycopene beta-cyclase (lcy, from daffodil). Scientists also

inserted other pieces of DNA that the genes needed to function in the cell, and they inserted marker

genes to help them track the inserted DNA. Then the scientists grew, selected, and tested the

embryos for beta-carotene. When full-grown, the rice plants produced and stored beta-carotene in

their starch.
3. ENHANCED TRAIT: Dolly the Sheep (what made Dolly so special
was that she had been made from an adult cell, which no-one at
the time thought was possible.)
MODIFYING TECHNIQUE: Cloning
Dolly was important because she was the

first mammal to be cloned from an adult

cell. Her birth proved that specialised cells

could be used to create an exact copy of

the animal they came from. This knowledge

changed what scientists thought was

Dolly and her
possible and opened up a lot of possibilities
surrogate mother
in biology and medicine, including the

development of personalised stem cells

known as iPS cells.

References: https://www.history.com/this-day-in-history/first-successful-

cloning-of-a-mammal

https://askabiologist.asu.edu/content/story-dolly
Dolly was part of a series of experiments at The Roslin Institute that were trying to develop a better

method for producing genetically modified livestock. If successful, this would mean fewer animals

would need to be used in future experiments. Scientists at Roslin also wanted to learn more about

how cells change during development and whether a specialised cell, such as a skin or brain cell,

could be used to make a whole new animal.

These experiments were carried out at The Roslin Institute by a team led by Professor Sir Ian Wilmut.

Because of the nature of the research, the team was made up of many different people, including

scientists, embryologists, surgeons, vets and farm staff.

Dolly was cloned from a cell taken from the mammary gland of a six-year-old Finn Dorset sheep and

an egg cell taken from a Scottish Blackface sheep. She was born to her Scottish Blackface

surrogate mother on 5th July 1996. Dolly’s white face was one of the first signs that she was a clone

because if she was genetically related to her surrogate mother, she would have had a black face.

Because Dolly’s DNA came from a mammary gland cell, she was named after the country singer

Dolly Parton
4. ENHANCED TRAIT: Genetically Modified Cows (tender, healthier --
similar to wagyu beef)
MODIFYING TECHNIQUE: Cloning
Professor Ni Minhong, who led the research at Beijing

University of Agriculture's department of advanced science

and technology, said it was "a crucial step".

"Through this project we will be the first in the world to

successfully create transgenic cows with fatty acid binding

protein," he said.

"Unlike pork where leaner is better, a good amount of

muscle fat content is one of the key elements when it

comes to characterising beef quality.

"After more research it may be possible to achieve ideal

marbling of meat in domestic cattle and provide an

alternative to imported high-grade meat."

Jing Qin 1 and 2 are a local Chinese breed of cattle called Qinchuan, but

have had a gene which encourages the creation of what is called the

adiposcyte fatty acid binding protein – the protein which creates a high

number of thin streaks of fat between muscles.

The fat, which becomes marbling after the animals have been slaughtered,

makes meat tender and creates its flavour.

John Torode, the MasterChef judge and author a book called "Beef", said

marbling is the key difference between a steak with flavour and one

without/].

"The strands of fat within the muscle work like a network and when cooked

this fat melts. The melting fat soaks into the fibres of the muscles making it

more tender and moist," he said.

"If you have a muscle that is long and lean, with little fat, it will just be tough once cooked.

Having that internal network of fat deposits in the muscle makes all the difference then

between dry, stringy meat and sumptuous, tender meat."

Wagyu beef has become famous around the world for its tenderness, and is the most

expensive beef available. It is often known as Kobe beef because of its production in the

area of Japan, but is now bred in Britain for sale in British restaurants.

Allowing genetically modified cattle would cut the cost of richly marbled beef.

Tests on the two calves have shown that they both carry the gene but it will be some

months before further tests can be conducted to find out if their muscles are becoming

enriched with fat.

Reference: https://www.businessinsider.com/researchers-tinker-with-

cow-genetics-to-make-tastier-beef-2012-8
5. ENHANCED TRAIT: (HoneySweet’ (C5), the First Genetically Engineered
Plum pox virus–resistant Plum
MODIFYING TECHNIQUE: Recombinant DNA Technology
‘HoneySweet’ originated as a seedling from the open pollination of ‘Bluebyrd’ plum (Scorza and Fogle,

1999). The pollen parent of ‘HoneySweet’ is unknown. ‘HoneySweet’ was originally selected in vitro as a

regenerated shoot from a hypocotyl slice that had been transfected with Agrobacterium tumefaciens

EHA 101 carrying the plasmid pGA482GG/PPV-CP-33 (Scorza et al., 1994). The regenerated,

transgenic shoot, coded as C5, along with other transgenic shoots, was rooted in vitro and transferred

to a greenhouse. Following greenhouse testing using graft and aphid inoculations with the M and D

strains of Plum pox virus (PPV), C5 (later patented as ‘HoneySweet’), was asexually propagated by bud

grafting on to standard rootstocks, including Prunus persica (GF305 peach seedlings), Prunus

domestica (European plum seedlings), Prunus myrobalan, and GF 8-1 (Prunus cerasifera × P.

munsoniana)

Reference: https://www.businessinsider.com/researchers-tinker-with-

cow-genetics-to-make-tastier-beef-2012-8
‘HoneySweet’ is heterozygous for the PPV-CP insert; therefore ≈ 50% of the male and female gametes leading to pollen and

egg development will contain the PPV-resistance transgene. When used as a parent in conventional hybridization,

≈
‘HoneySweet’ transfers the PPV-CP transgene insert to progeny as a single dominant locus and, as predicted by Mendelian

genetics, 50% of the progeny carry the transgene insert. These are resistant to PPV (Ravelonandro et al., 2002; Scorza et

al., 1998), making ‘HoneySweet’ a useful source of resistance for developing additional PPV-resistant cultivars. ‘HoneySweet’

is also highly resistant to black knot disease caused by the fungus Apiosporina morbosa. It appears to have inherited this

resistance from ‘Bluebyrd’, its seed parent, which is also highly resistant to black knot (Scorza and Demuth, 2015).

PPV Resistance

Sharka disease caused by PPV affects most stone fruit species, cultivated and wild (Damsteegt et al., 2007). Symptoms of

Sharka disease are described in Levy et al. (2000). Although resistance to PPV has been pursued ever since the disease was

first identified, there are few reports of high-level resistance in commercial Prunus species and new sources of high-level

resistance to PPV are needed.

Initial ‘HoneySweet’ PPV-resistance tests were greenhouse-based (Ravelonandro et al., 1997). Although no PPV field

inoculations were performed in the United States due to the quarantine status of PPV, through nearly 20 years of field

testing in Europe, under heavy natural aphid-vectored infection pressure, of the 99 trees of ‘HoneySweet’ that have been

evaluated, none have become infected with PPV through natural aphid transmission, whereas control trees have become

rapidly and severely infected.

Fig. 1.

(A) ‘HoneySweet’ plum (left);

‘HoneySweet’ plum harvest at the

U.S. Department of Agriculture–

Agricultural Research Service,

Kearneysville, WV (right). (B) Three-

year old ‘HoneySweet’ tree in

Bistrita, Romania (left), with a

weighed sample of ‘HoneySweet’

fruit (right). Photo credits: Scott

Bauer, Peggy Greb, and Ioan

Zagrai.