GENERAL BIOLOGY 2

Governor Pack Road, Baguio City, Philippines 2600

Tel. Nos.: (+6374) 442-3316, 442-8220; 444-2786;
442-2564; 442-8219; 442-8256; Fax No.: 442-6268 Second Trimester
Email: email@[Link]; Website: [Link]

MODULE 1 – Gen Bio 2 Subject Teacher:

Outline:
a. Genetic engineering vs Classical breeding
b. Tools used in Recombinant DNA technology
c. Methods of Gene Cloning for DNA Recombination
d. Transgenic organisms
e. Classical genetics

Learning Objectives:
After completing the module, the students will be able to:
1. Outline the processes involved in genetic engineering;
2. b. Discuss the applications of recombinant DNA;
3. c. Discuss Mendelian genetics.

GENETICS (Part 1)

INTRODUCTION:

The DNA and Genetic Information Nucleic acids are complex molecules containing C, H, O, P and
N elements. They are composed of polynucleotide chains. There are two types of nucleic acids:
DNA and RNA. The DNA is responsible for storing information that serve as basis for synthesis of
proteins that in turn control all reactions and formation of structures. The RNA plays several roles in
the actual building of proteins. (Review the structure and functions of nucleic acids as well as the
process of protein synthesis in General Biology 1 notes if necessary)

MODERN GENETICS: RECOMBINANT DNA TECHNOLOGY

Genetic engineering or recombinant DNA technology refers to various techniques and procedures
used in gene manipulation. However, natural DNA recombination occurs randomly during the
crossing-over of homologous chromosomes during Meiosis I (review stages of Meiosis), during
fertilization when two gametes fuse, or even through mutation. DNA recombination, both natural and
technology-manipulated, is being used to advantage in improving production of agricultural
products, forensics, pharmaceuticals and medicine.

Modern Genetic Techniques vs Classical Breeding

Table 8.1 Genetic engineering compared to classical breeding

Genetic Engineering Classical Breeding

It involves the use of molecular techniques to It focuses on mating of organisms with desirable
modify the traits of target organisms. These are qualities. This relies heavily on the naturally
often referred to as transgenic organisms or occurring life cycle of organisms and
genetically modified organisms (GMO). homologous recombination to eliminate
undesirable traits.
Genetic modification is achieved by adding a Interbreeding can only be carried out with
specific gene or genes or removing undesirable closely or distantly related organisms.
gene, to produce a desirable phenotype. The
gene can be from the same species or different
organism.
Sometimes, the desired trait is attained faster It is time-consuming as breeding requires series of
than classical breeding. crossing and self-fertilization to be able to attain
desired trait.
 GENERAL BIOLOGY 2
Governor Pack Road, Baguio City, Philippines 2600
Tel. Nos.: (+6374) 442-3316, 442-8220; 444-2786;
442-2564; 442-8219; 442-8256; Fax No.: 442-6268 Second Trimester
Email: email@[Link]; Website: [Link]

MODULE 1 – Gen Bio 2 Subject Teacher:

Tools Used in Recombinant DNA Technology

• Target DNA- This is the gene of interest, the gene sequence that codes for the desired trait.
• Restriction enzymes- These enzymes identify specific base sequence and cut the DNA at a
specific point between two nucleotides.
• DNA cloning vectors- These are DNA materials used to carry the target DNA to be transferred.
Examples are bacterial plasmids and bacteriophages (bacteria-infecting viruses).
• Host cell- A cell that is able to accept the target DNA to multiply or replicate it. Host cells must
be able to maintain the structure of the recombinant DNA all throughout generations.
• Modifying enzymes- These are enzymes involved in degradation, synthesis and alteration of
the nucleic acids. Examples are DNA ligase and Taq Polymerase.

Methods of Gene Cloning for DNA Recombination

The process of gene cloning involves a complex process mostly varied but almost similar with
the concepts of natural processes of DNA replication and protein synthesis. But to simplify the entire
process, there are four stages of gene cloning:

1. Isolation of target Gene- Cutting the gene from the original chromosome or producing a
complementary DNA by replicating the target gene.
2. Insertion of target gene into the vector- The target gene is inserted into the vectors resulting
to recombinant DNAs.
3. Introduction of vector to the host cell
4. Amplification of the target gene by the host cell- The host cells are allowed to reproduce to
produce multiple copies of the target genes.
 GENERAL BIOLOGY 2
Governor Pack Road, Baguio City, Philippines 2600
Tel. Nos.: (+6374) 442-3316, 442-8220; 444-2786;
442-2564; 442-8219; 442-8256; Fax No.: 442-6268 Second Trimester
Email: email@[Link]; Website: [Link]

MODULE 1 – Gen Bio 2 Subject Teacher:

Some Examples of Transgenic Organisms

1. BT Corn- Gene sequences from Bacillus thuringiensis that code for the production of protein
that is toxic to corn pests.
2. Golden Rice-Scientists are working on integrating genes to rice so it can synthesize betacarotene
3. Transgenic Atlantic salmon- modified genetically to enhance growth rate
4. Transgenic Ruminants- genetically enhanced to produce milk with pharmaceutical proteins
5. Human insulin producing E. coli- Gene sequence from human for production of insulin was
inserted in E. coli bacteria giving the bacteria the ability to produce human insulin.
6. Genetically modified Pseudomonas bacteria- The bacteria were modified to enhance their
ability to degrade hydrocarbons so they can be used in cleaning up oil spills.

CLASSICAL GENETICS: MENDELIAN GENETICS

Definition of Terms
• Gene- the sequence of DNA that codes for a specific trait. E.g. The DNA sequence that codes
for the color of the eyes. These are represented by any letter in genetic problems.
• Allele- the alternative forms of genes (minimum of two). E.g. The color of the eyes differ among
individuals. Blue and brown are the most common. The genes for these are known as alleles.
• Locus- the location of the genes in the chromosomes
• Dominant- The allele that is expressed in case more than one allele is present. These are
represented by UPPERCASE letters. Example, in a pair of chromosomes, one gene codes for
blue eyes and the other is for brown eyes. The appearance that will be expressed is brown
eyes. This means that brown is dominant over blue. Therefore, if we are to represent using the
letter B, B is for brown and b is for blue.
• Recessive- the allele that is not expressed in case more than one allele is present. These are
represented by lowercase letters. In the given example above, the gene for blue eyes is
recessive. Therefore, even if it is present, if the gene for brown eyes is present, it will not be
expressed. It will only be expressed if both alleles present are for blue eyes.
• Homozygote- a combination of similar alleles. In the example given above, if the combinations
are both genes for brown eyes (BB) or both genes for blue eyes (bb), they are homozygote:
BB-homozygous dominant; bb-homozygous recessive
• Heterozygote- combination of different alleles. In the example given above, a combination of
a gene for brown eyes and a gene for blue eyes (Bb) is known as heterozygote.
• Genotype- is the combination of genes represented by letters. They are always written in pairs.
Examples: BB, Bb, bb
• Phenotype- the appearance of the specified genotype. Examples: BB-phenotype: Brown eyes,
Bb- phenotype: Brown eyes, bb-phenotype: blue eyes
• Test cross- These are used to determine if an individual with dominant trait is homozygote or
heterozygote. The individual with unknown genotype is crossed to a homozygous recessive.
• Parental Generation (P)- the first genotypes that are crossed; First filial generation (F1)-
generation after P; Second filial generation (F2)- generation after F1.

Mendelian Laws
Gregor Mendel, an Austrian monk, is known as the Father of Genetics due to his discoveries on
the passing of traits from one generation to another. His famous garden peas experiment
revealed the concepts of heredity though during his time, there are no concepts of nucleic acids
and chromosomes. Through his experiments, he came up with the following laws:

A. Law of Segregation- Alleles coming from the parents combine during fertilization. Therefore,
there are two alleles per trait in an individual. However, during gamete formation (Meiosis),
these pair of alleles will separate or segregate. (Take note, in meiosis, diploid cells will produce
haploid daughter cells). Therefore, when they are passed to the next generation, either but
not both of the alleles are passed to one offspring. Let’s explain this using monohybrid crosses
or crosses involving one trait only.
 GENERAL BIOLOGY 2
Governor Pack Road, Baguio City, Philippines 2600
Tel. Nos.: (+6374) 442-3316, 442-8220; 444-2786;
442-2564; 442-8219; 442-8256; Fax No.: 442-6268 Second Trimester
Email: email@[Link]; Website: [Link]

MODULE 1 – Gen Bio 2 Subject Teacher:

Examples:
1) Let us use one trait observed by Mendel on color of the seed of peas.
Y-yellow (dominant), y-green (recessive)
Let’s say a plant with yellow seed (heterozygous) is allowed to self-pollinate. What will be the
seed color of the offspring?
Answer: First, we have to assign the genotypes of the parents based from the problem. So, it
says that a heterozygous was allowed to self-pollinate. Dominant trait is yellow seed color, and
heterozygous means both alleles for seed color, yellow (Y) and green (y), are present.
Therefore, the genotypes of P (parental generation) will be both Yy. According to the Law of
Segregation, these alleles will segregate or separate during meiosis (gamete formation).
Therefore, there are two types of genes that will be contained in the gametes that will be formed.
Some gametes will contain the gene for yellow seed color (Y), and some will contain
the gene for green seed color (y). Therefore, if the plant will self-pollinate, then they will
produce offspring with phenotypes and genotypes as shown in the Punnet Square below:

P(Parental Generation): Yy x Yy (Both alleles will segregate forming Y and y, and another Y
and y. Note, this is self-pollination, therefore, the parents have the same genotype)

Punnet Square
Stamen (male) The highlighted boxes represent the possible combination of
Y y alleles of the offspring based from the segregated alleles of the
(female)

Y YY Yy parents. Therefore, there are 3 possible genotypes of the

offspring or the first filial generation (F1). These are YY, Yy and yy
Pistil

y Yy yy
as seen on the Punnet Square.

2) A pure breed/ homozygous garden pea with purple flower was cross pollinated to a garden
pea with white flower. Mendel’s experiment concluded that purple flowered are dominant
over white. What will be the genotypic and phenotypic ratio of the offspring?

Answer: (Let’s assign P as purple and p as white)

P: Parent 1 is homozygous dominant so the genotype is PP. Parent 2 is homozygous recessive;
therefore, the genotype is pp.

Punnet Square
Parent 1 F1:
P p Pp=purple
Parent 2

P PP Pp GR= 4 Pp
p Pp pp PR= 4 purple or 100% purple

B. Law of Independent Assortment- Separate genes for separate traits are passed independently
of one another. Genes do not influence each other with regards to the sorting of alleles into
gametes. Therefore, combination of alleles for every gene is equally likely to occur. These are
best explained in dihybrid or polyhybrid crosses (involving two or more traits).
 GENERAL BIOLOGY 2
Governor Pack Road, Baguio City, Philippines 2600
Tel. Nos.: (+6374) 442-3316, 442-8220; 444-2786;
442-2564; 442-8219; 442-8256; Fax No.: 442-6268 Second Trimester
Email: email@[Link]; Website: [Link]

MODULE 1 – Gen Bio 2 Subject Teacher:

Examples:
1) In garden peas, round seed shape is dominant over wrinkled. A garden pea has round,
yellow seed. It was pre-determined that it is heterozygous for both traits. It was allowed to self-
pollinate. What will be the genotypic and phenotypic ratio of offspring?

P: Let’s assign R for round and r for wrinkled and retain Y for yellow and y for green seed color.
Based from this, since it is heterozygous for both traits, then the genotype is Rr for seed shape
and Yy for seed color. We are going to combine them so the genotype for both parents will
be RrYy. Each pair will segregate, forming R and r, then Y and y. Now, according to the law of
independent assortment, these genes will be passed to offspring independently. Meaning,
they will not affect the distribution of each other. Therefore, there are 4 possible combinations
during segregation. These are: RY, Ry, rY and ry. Now that the combinations in the gametes
were determined, we proceed to the Punnet Square.

2) A garden pea that is heterozygous for flower color (purple) with wrinkled seed shape is crossed
with another garden pea which is heterozygous for both flower color and seed shape (round).
What is the genotypic and phenotypic ratio of the offspring?

P: Let’s assign the genotypes. Parent 1 is heterozygous for purple (Pp) with wrinkled seed (rr). So
parent 1 has a genotype of Pprr. Parent 2 is heterozygous for both flower color (Pp) and seed
shape (Rr). Therefore, the genotype is PpRr. For parent 1, there are 2 types of alleles for flower
color, P and p but only one type of allele for seed shape (r). Therefore, there are only 2
possible combinations of alleles in gametes: Pr and pr. On the other hand, for parent 2, there
are 2 types of alleles for flower color, P and p. Also, there are 2 types of alleles for seed shape,
R and r. Therefore, for parent 2, there will be 4 possible combinations of alleles in the gametes:
PR, Pr, pR and pr.
Parent 1: Pr, pr
Parent 2: PR, Pr, pR, pr

F1:
GR: 1 PPRr: 2 PpRr: 1 PPrr: 2 Pprr: 1ppRr: 1 pprr (or simply 1:2:1:2:1:1)
PR: 3 purple and round: 3 purple and wrinkled: 1 white and round: 1 white and wrinkled (3:3:1:1)