APPLICATIONS IN

BASIC RESEARCH
(IDENTIFY, MAP, CLONE, AND
SEQUENCE GENES AND TO
DETERMINE THEIR FUNCTIONS)

PRESENTED BY- PRESENTED TO -

MEGHA DR MANDEEP
9052 DEPARTMENT OF BOTANY
BSC HONS ZOOOGY HANSRAJ COLLEGE
 INTRODUCTION

BASIC RESEARCH - It focuses on exploring the structure, function, and regulation of

genes, which are the fundamental units of heredity. This field investigates how genes are
organized within the genome, how they encode proteins, and how they interact with one
another and with the environment. By delving into the molecular mechanisms of
genetics, researchers aim to uncover the principles governing heredity, variation, and
biological processes across different organisms.
 GENE IDENTIFICATION

Gene identification is the process of determining the location and

function of specific genes on chromosomes. This crucial step in
genetics enables researchers to link genetic information with
traits and diseases, providing a foundation for understanding
complex biological systems.

Methodologies
Several methodologies are employed in gene identification:

Genetic Markers: These are specific DNA sequences that can indicate the presence of a
particular gene. Researchers use markers to track inheritance patterns and associate
them with specific traits or diseases.
Comparative Genomics: This approach involves comparing the genomes of different
species to identify conserved genes and genomic regions. By examining similarities and
differences, scientists can infer the function of genes and their evolutionary
significance.
Phenotypic Analysis: This method assesses the physical traits (phenotypes) of
organisms in relation to their genetic makeup. By correlating observable characteristics
with genetic data, researchers can pinpoint genes that influence specific traits.
 GENE IDENTIFICATION

Applications
Gene identification has significant applications in various fields:
Disease Association: By identifying genes linked to diseases, researchers can better understand
the genetic factors contributing to conditions like cancer, cystic fibrosis, and heart disease. This
knowledge is vital for developing targeted therapies and personalized medicine.
Genetic Counseling: Understanding gene functions and their associations with hereditary
diseases allows for better risk assessment and counseling for individuals and families affected by
genetic disorders.

Example
A well-known example of gene identification is the BRCA1 and BRCA2 genes, which are
linked to an increased risk of breast and ovarian cancer. Mutations in these genes can
significantly raise the likelihood of developing these cancers, leading to the development
of genetic tests that help in assessing risk and guiding preventive measures. Identifying
these genes has transformed approaches to cancer screening and has informed
treatment strategies, exemplifying the critical role of gene identification in healthcare.
 Types of Gene Maps:
GENE MAPPING
1. Genetic Maps (Linkage Maps):
These maps are created by
studying the recombination Gene Mapping is the process of determining
frequency between genes during the relative positions of genes on a
meiosis. Genes that are closer chromosome. It helps in understanding the
together on a chromosome tend genetic architecture of organisms, including
to be inherited together, and this plants, animals, and humans
information is used to estimate
their positions relative to each
other. Techniques Used for Gene Mapping:
2. Physical Maps: Linkage Analysis:
These maps are based on the This traditional method uses recombination data from breeding
experiments or family studies to estimate the relative positions of genes.
actual physical distance between Genome-Wide Association Studies (GWAS):
genes, measured in base pairs GWAS is a more recent approach that looks at genetic variations across the
entire genome to identify loci associated with specific traits or diseases,
(bp). Techniques like DNA based on statistical correlations in large populations.
sequencing are used to create Fluorescence In Situ Hybridization (FISH):
FISH is a technique that uses fluorescent probes to bind to specific DNA
high-resolution physical maps. sequences on chromosomes, allowing scientists to visually locate genes
under a microscope.
Applications of Gene Mapping:
GENE MAPPING
Identifying Genetic Loci:
Gene mapping is crucial in Example :
identifying genetic loci In tomato plants, gene mapping has been used to identify genes
responsible for traits such as responsible for resistance to pathogens like Phytophthora
disease resistance, flowering infestans (which causes late blight). Linkage maps helped
time, or fruit size in plants. researchers to pinpoint resistance genes (R genes), while
Understanding Diseases: physical maps provided precise locations, enabling plant
In medicine, gene mapping breeders to develop more resistant varieties.
helps in pinpointing genes
associated with hereditary
diseases, aiding in
diagnostics and therapeutic
developments.

GENE MAPPING
 GENE CLONING

Gene cloning is the process of creating copies of specific genes or DNA segments for the purposes of
analysis, manipulation, and study. This technique allows scientists to isolate and replicate genes,
providing a deeper understanding of their function and behavior.

Techniques Used in Gene Cloning:

1. Recombinant DNA Technology:

This involves combining DNA from different sources. A gene of interest is inserted into a vector (usually
a plasmid), which is then introduced into a host cell (such as bacteria or yeast) to replicate the gene.
2. Plasmid Vectors:
Plasmids are small, circular pieces of DNA found in bacteria. They are commonly used as vectors to
transfer foreign DNA into host cells. Plasmids can carry genes of interest along with selectable markers,
enabling researchers to identify successfully transformed cells.
3. CRISPR-Cas9:
This is a revolutionary gene-editing technology that allows for precise modifications of DNA. It uses a
guide RNA to direct the Cas9 enzyme to a specific location in the genome, where it can cut the DNA,
allowing for the insertion or deletion of genes.
 GENE CLONING

Example
Applications in Research: Development of Bt cotton. In this case, the gene
from the bacterium Bacillus thuringiensis (Bt) that
Gene cloning has numerous applications :
Study of Gene Expression:
produces a protein toxic to certain pests (like the
Researchers can clone genes to analyze their expression cotton bollworm) was cloned and inserted into the
patterns in different conditions, such as during stress or cotton plant's genome
development. This helps in understanding how plants
respond to environmental changes.
Protein Production:
Cloned genes can be used to produce specific proteins,
such as enzymes or hormones, which can be studied for
their roles in plant physiology or used in biotechnological
applications.
Creation of Genetically Modified Organisms (GMOs):
Gene cloning enables the introduction of desirable traits
into plants, such as pest resistance or increased nutritional
value. This is achieved by inserting specific genes into the
plant genome, leading to improved crop varieties.

BT COTTON
 GENE SEQUENCING
Technologies Used: Gene sequencing refers to determining the
exact order of nucleotides (A, T, C, G) in a
1. Sanger Sequencing:
A traditional method where short DNA fragments are
gene or an entire genome.
sequenced individually. While accurate, it is slow and
costly for large genomes.
2. Next-Generation Sequencing (NGS):
A more advanced, high-throughput method that allows
rapid sequencing of entire genomes or large numbers
of genes simultaneously. NGS is widely used for plant
research due to its efficiency and scalability.
3. CRISPR-based Approaches:
CRISPR technology is used not only for gene editing
but also for targeted sequencing. It can precisely
identify and sequence specific regions of DNA,
enabling researchers to study functional genes in
detail. SANGER SEQUENCING
 Applications :
GENE SEQUENCING Genome-Wide Association Studies (GWAS):
Sequencing plant genomes helps identify
genetic variations associated with important
traits like disease resistance, drought
tolerance, or yield. GWAS links specific genes
with phenotypic traits across populations.
EXAMPLE Precision Agriculture:
Sequencing of the rice genome. By sequencing the Sequencing allows for the development of
crops tailored to specific environments,
genome of Oryza sativa (rice), researchers identified improving resistance to pests, diseases, and
key genes responsible for traits like drought tolerance environmental stressors. This enables more
and pest resistance. This knowledge has enabled the targeted plant breeding programs and the
creation of genetically modified organisms
development of improved rice varieties, particularly (GMOs).
important for food security in regions that rely on rice Evolutionary Studies:
Genome sequencing provides insights into
as a staple crop. evolution, speciation, and domestication. By
comparing genomes of different plant species,
scientists can track how plants have adapted
to different environments over time.
 Approaches to Functional Analysis:
FUNCTIONAL 1. Gene Knockout:

ANALYSIS OF GENE Involves disabling or "knocking out" a

gene to observe what happens in its
absence. This helps in identifying the
gene's function by noting changes in the
organism's phenotype or behavior.
2. Gene Overexpression:
The process of increasing the expression
Functional analysis refers to of a particular gene to study its
understanding the role and biological enhanced effects. Overexpression can
function of genes in cells and organisms reveal how higher levels of a gene's
product (protein) affect the organism's
traits.
3. RNA Interference (RNAi):
A method where small RNA molecules
are used to silence or reduce the
expression of specific genes. RNAi is
useful for knocking down gene
expression temporarily, allowing
researchers to study the gene's function
without a permanent knockout.

RNA INTERFERANCE
 FUNCTIONAL
Applications:
Uncovering Gene Function:
ANALYSIS OF GENE
Functional analysis helps in identifying the
roles of genes in plant growth,
development, and responses to
environmental stress.
Developing GMOs: EXAMPLE
Genes that enhance resistance to pests, Study of the DREB1A gene in Arabidopsis.
diseases, or harsh conditions can be studied Researchers used gene overexpression to introduce
and then modified for crop improvement. the DREB1A gene into plants like rice and wheat.
The DREB1A gene is known to be involved in
Studying Cellular Pathways:
regulating drought tolerance. Overexpression of
By understanding the function of specific this gene resulted in improved drought tolerance in
genes, researchers can map out cellular these crops. Conversely, gene knockout studies in
pathways involved in processes like Arabidopsis showed that when DREB1A was
photosynthesis, nutrient uptake, or disabled, the plants were more susceptible to
hormonal signaling. drought stress.
 CONCLUSION

In conclusion, the applications of molecular tools in gene identification,

mapping, cloning, sequencing, and functional analysis are transformative.
These advanced techniques enable researchers to gain deeper insights into
plant genetics, facilitating the discovery of genes responsible for essential
traits such as drought tolerance, disease resistance, and nutrient efficiency.

As we continue to refine these methodologies, we unlock the potential to

improve agricultural practices significantly, leading to higher yields and
more resilient crops in the face of climate change and food security
challenges. The integration of molecular biology into plant research not only
enhances our understanding of plant biology but also holds great promise
for developing innovative solutions that address global challenges related to
food production and environmental sustainability.
THANK YOU