MOLECULAR BIOLOGY

(ZEB 361)

BIOLOGICAL MOLECULES

Biological molecules include Carbohydrates, Lipids, Proteins, Nucleic Acids,

Enzymes, Vitamins, Hormones, Cholesterol, Amino Acids, Phospholipids. Much of
the cell content is an aggregation of small molecules (e.g., simple sugars, amino
acids, vitamins) and ions (e.g., sodium, chloride, calcium ions). The locations
and concentrations of small molecules and ions within the cell are controlled by
numerous proteins inserted in cellular membranes. Certain small molecules
(monomers) in the cell can be joined to form polymers through repetition of a
single type of chemical-linkage reaction (bonding).

Cells produce three types of large polymers, commonly called macromolecules

namely polysaccharides, proteins, and nucleic acids. Sugars, for example, are
the monomers used to form polysaccharides.

NUCLEIC ACIDS

The principal genetic molecules of living organisms are chemically called nucleic
acids. Both DNA and RNA are polymers of complex molecules containing carbon,
oxygen, hydrogen, nitrogen and phosphorus. The molecules of nucleic acids
(polymers) are composed of monomers called nucleotides joined by covalent
bonds.

Nucleic acids are extremely complex molecules which help to pass on hereditary
characteristics from one generation to the next, and to trigger the manufacture
of specific proteins. Nucleic acid molecules are very large chains of repeating
nucleotide units linked in many sequences. Thus, nucleic acids are high
polymers with very high molecular weights.

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A nucleotide is a molecular unit of a nucleic acid molecule that consists
of 3 subunits namely, a phosphate group, a pentose sugar (ribose or
deoxyribose) and a nitrogen base (purine or pyrimidine).

A nucleoside is a compound consisting of 2 subunits – a pentose sugar and a

nitrogen base. It is a precursor of a nucleotide. A summary of nucleic acid
formation is:

· Pentose sugar + Nitrogen base = Nucleoside

· Nucleoside + Phosphate group = Nucleotide

· Nucleotide + Nucleotide + Nucleotide = Nucleic acid.

Types of nucleic acids

There are two types of nucleic acids – DNA (Deoxyribonucleic acid) and RNA
(Ribonucleic acid); both are chemical relatives that are universally present in all
living cells, and they form the chemical basis of life. Both DNA and RNA contain
the purines – Adenine (A) and Guanine (G) and the pyrimidine cytosine (C). The
second kind of pyrimidine in DNA is Thymine (T) whereas it is Uracil (U) in RNA.
Therefore, this unique pyrimidine distinguishes DNA from RNA. DNA is a double-
stranded molecule that stores genetic information, while RNA is a single-
stranded molecule that is involved in the synthesis of proteins.

- DNA

The DNA is a polymer made up of repeating units of mononucleotides carrying

the genetic material of all cellular organisms. DNA carries the information
needed to direct protein synthesis and replication. Protein synthesis is the
production of the proteins needed by the cell for its activities and development.
Replication is the process of which DNA copies itself for each descendant cell,
passing on the information needed for protein synthesis. In most cellular
organisms, DNA is organised on chromosomes located in the nucleus of the cell.

- RNA

Ribonucleic acid (RNA) in cellular organisms is the molecule that directs the
middle steps of protein production. In cellular organisms the DNA, carries the

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information that determines protein structure, but DNA cannot act alone and
relies upon RNA to transfer this crucial information (translate) during protein
synthesis – production of the proteins needed by the cell for its activities and
development. RNA differs chemically from DNA by being single stranded, having
a ribose sugar instead of Deoxyribose sugar and having uracil as nitrogenous
base instead of thymine.

There are three types of RNA classified based on their molecular size. The
smallest type of RNA is called transfer – RNA (tRNA) which carries amino acids
to the ribosomes for incorporation into a protein. Each amino acid has different
classes of tRNA that read the codes of mRNA, therefore involved in protein
synthesis. The tRNA receives information from mRNA, through pairing of their
bases and accordingly selects specific amino acids and pass to the ribosome.

The second type of RNA is the ribosomal – RNA (rRNA), this is larger than tRNA
and composes the ribosomes in the cytoplasm, the specialised structures that
are the sites of protein synthesis.

The largest type of RNA is the messenger – RNA (mRNA). Messenger – RNA is
a strand of RNA that is complementary to the DNA sequence for a gene
and carries the genetic blueprint copied from the sequence of bases in a
cell’s DNA. This blueprint specifies the sequence of amino acids in a protein.

PROTEIN

Proteins are macromolecules consisting of amino acids. The varied structures of

proteins enable them to carry out numerous functions. Amino acid is obtained
either by synthesizing them from other molecules or by breaking down proteins
that we eat. The “essential” amino acids, from a dietary standpoint, are those
we cannot synthesize and must obtain from food.

Proteins can serve as structural components of a cell, for example, by forming

an internal skeleton (hair). They can be sensors that change shape as
temperature, ion concentrations, or other properties of the cell change
(hormones). They can import and export substances across the plasma
membrane (haemoglobin). They can be enzymes, (lactase) causing

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chemical reactions to occur much more rapidly than they would without
the aid of these protein catalysts. They can be extracellular signals,
released from one cell to communicate with other cells, or intracellular signals,
carrying information within the cell (insulin).

Other examples of proteins include antibodies, collagen, elastin, keratins.

Structure

Cells string together 20 different amino acids in a linear chain to form a protein.
The structure of amino acids consists of four groups attached to a central carbon
atom: a hydrogen atom, an amino group, a carboxyl group, and a side chain or
R group. Amino acids are linked together by their amino and carboxyl groups.
The R group (reactive groups) or side chain gives the protein its distinguishing
properties and functional specificity.

When amino acids polymerize, the carboxyl group of one amino acid reacts with
the amino group of another to form a peptide linkage. A polymer of amino acids
connected by peptide linkages is a polypeptide. A protein is made up of one or
more polypeptides. At both ends of the polypeptide is a free amino and carboxyl
group. The primary structure of a protein is determined by the gene
corresponding to the protein. A specific sequence of nucleotides in DNA is
transcribed into mRNA, which is read by the ribosome in a process called
translation.

Elevated temperatures, pH changes, or altered salt concentrations can disrupt

the structure of a protein (denaturation). Increased temperature can break weak
hydrogen bonds. Altered pH can change the pattern of ionization of carboxyl and
amino groups in the side chains of amino acids.

RECOMBINANT DNA TECHNOLOGY

Recombinant DNA (rDNA) technology is a process in which DNA from two

different sources is combined to create a new, recombined DNA molecule. This

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technology is used to create new proteins, modify existing proteins, and even
create new organisms. The applications of recombinant DNA technology range
from creating transgenic plants and animals to producing medicines and
vaccines. It is also used in genetic engineering, where genes from one organism
are inserted into another organism to create a desired trait. The rDNA process
involves isolating a gene from one organism, cloning it into a vector (plasmids,
yeast, bacteria, viruses), and inserting it into another organism. This process
enables scientists to create new proteins and organisms that can be used to
benefit society. The benefits of rDNA technology include the ability to create new
medicines, crops, and other products that can improve human health and the
environment. Additionally, rDNA technology has been used to develop new
treatments for diseases such as cancer and AIDS.

Types of Recombinant DNA technology

1. Plasmid-Based Recombinant DNA Technology: This technology involves

the manipulation of plasmids, which are self-replicating, extrachromosomal
circular DNA molecules, to introduce foreign genetic material into a host cell.

2. Retroviral Vector Technology: This technology utilizes viral vectors derived

from retroviruses to introduce foreign genetic material into the host cell.

3. Polymerase Chain Reaction (PCR): This technique uses DNA polymerase to

replicate a specific region of DNA and amplify it for further analysis.

4. Restriction Fragment Length Polymorphism (RFLP): This technique uses

restriction enzymes to cut DNA into fragments, which are then separated and
studied to identify polymorphisms.

5. Site-Directed Mutagenesis: This technique is used to introduce mutations into

a specific region of DNA.

6. DNA Cloning: This technique involves copying and inserting a gene into a
vector such as a plasmid or bacteriophage, which is then used to produce large
amounts of the gene.

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7. Transgenic Technology: This technique involves transferring a gene from one
organism to another, resulting in the production of an organism containing the
new gene.

8. Gene Expression: This technique involves introducing a gene into a host

organism, which then expresses the gene, resulting in the production of the
protein encoded by the gene.

9. DNA Sequencing: This technique is used to determine the nucleotide

sequence of a gene or genome.

Uses of Recombinant DNA technology

1. Creation of Human Insulin: Recombinant DNA technology is used to create

human insulin, which is an important treatment for diabetes. Human insulin is
produced by inserting the gene for human insulin into bacteria or yeast, which
then produce the insulin in large quantities.

2. Vaccine Production: Recombinant DNA technology is used to produce vaccines

for diseases such as hepatitis B, rabies, and HPV. The DNA of the virus is
inserted into a harmless virus, which then produces the antigens that cause the
body to produce antibodies against the disease.

3. Gene Therapy: Recombinant DNA technology is used in gene therapy, which is

a potential treatment for genetic diseases. The defective gene is replaced with a
healthy gene, which is inserted into the patient’s cells.

4. Diagnostics: Recombinant DNA technology is used in the development of

diagnostic tests for genetic diseases. The defective gene is identified and used as
a marker to diagnose the disease.

5. Genetically Modified Organisms: Recombinant DNA technology is used to

create genetically modified organisms (GMOs). These organisms have had their
DNA altered to make them resistant to pests, produce more food, or possess
other desired traits.

6. Forensic applications: Recombinant DNA technology is used in forensics to

identify individuals from biological samples such as blood, hair and semen.

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