Subject:

Cell and Molecular biology

Presented by:

Saira Kiran

Roll # BS5M23F24

Semester:

BS 5th Semester Morning

Submitted to:

Dr. Nadeem Sheikh

Genetic Engineering and DNA recombinant technology Genetic Engineering:

It is a technique used for deleting, inserting or changing the specific gene of an organism to get
product of our choice with the help of laboratory technology. This process involves to change the
single base pair and adding or deleting new segment by replacing previous segment.

Recombinant DNA technology:

It is the method to isolate or combine the desired DNA segments to form new gene with new
functions.

Tools:

To manipulate the DNA different tools and methods of recombinant technology are used.
These tools play crucial role in manipulation analysis and expression of genetic material
including advancement in genetic engineering and recombinant technology. These tools are
very important and perform specific function in changing or manipulation of DNA. These
are also vital in biotechnology field covering the areas of disease treatment improvement in
agriculture and biofuels

1. Restriction enzyme
2. DNA ligase
3. PCR(amplifying,copies)
4. Electrophoresis(separate)
5. Cloning vector(replictate foreign DNA in host )
6. Gel electrophoresis(separating DNA fragments based on size)

Enzymes:

Enzymes are very important in the fields of biotechnology and also crucial for genetic
engineering. These enzymes help in the manipulation of DNA. Some enzymes are given
below which are used in genetic engineering and recombinant DNA technology:

� Restriction enzyme uses to generate DNA fragment with sticky end enabling the
insertion of foreign DNA into vectors.

� DNA ligase is also known as Glue of DNA and used for joining the DNA during the
formation of recombinant molecules.

� DNA polymerase catalyze the synthesis of DNA.

� RNA polymerase catalyze the synthesis of RNA.

 Vectors:

A piece of DNA carries genetic material of DNA or we can say that carrying
sequence of DNA is known as vector.

Vectors are carriers for transforming genetic material for expression and
isolation. They have ability to replicate with in the host cell and have specific
sites where researchers can easily insert DNA of choice. These vectors vary in
size and choice of vector depend on the size of DNA.

There are various types of vectors like:

 Plasmid
 Bacteriophage cosmid vector,
 Bacterial artificial chromosomes
  Yeast artificial chromosome.

The most common is plasmid vector.

 Recombination of DNA:
The process of analyzing or combining the fragments of DNA from one or several
organisms to produce desired DNA through different genetic material. This
technology is called recombinant technology and this technology was firstly
introduced by Swiss microbiologist in 1968. The steps for recombinant technology
are given membrane or cell wall. it can be done by lysozymes for bacterial cells,
cellulose for plant cells and chitins for fungi cells and restriction endonuclease for
animal cells.

Selection of Recombinent:

It refers to the process by which two or molecules change genetic material

or combine to create new DNA sequences.This process is essential for diversity
and evolution occurs in various organisms

Here are common method for selecting recombinant :

Antibiotic resistence:
Plasmids used as vectors antibiotic resistence oftrn contain antibiotic
resistence genes.
Reporter resistence:
These are genes that code for a visible or measurable marker,such as the
green florescent protein (GFP).
If the foreign DNA is successfully inserted ,the host cell will express the reporter
gene,allowing for the identification of recombinants.
Color selection:
Some recombinant plasmids can produce colored colonies due to the
insertion of a DNA fragment .For example, the blue \white screening system is
often used with beta-galactosidase in E-coli for selecting recombinant colonies.

Replica plating:

1.Cultivating microorganism on primary agar plate:

● In the initial phase, cultivate the desired microorganisms on an agar plate. The
composition of this plate may include specific selective agents or lack a particular nutrient,
depending on the goals of the experiment.

2.Utilizing velvet impression replica technique:

● Employ a replica plating device covered with sterile velvet to create an impression on the
surface of the agar plate. This step captures a replica of the microbial colonies while
maintaining their original spatial arrangement.

3. Transferring Microbial Colonies to Secondary Agar Plates:

● Use the velvet replica to transfer the microbial colonies onto multiple
secondary agar plates. This process ensures the smooth relocation of the
microorganisms. 4. Screening for comprehensive analysis of the
experiment’s outcomes Phenotypic Traits: ● Conduct screenings to identify
specific phenotypic traits as part of a comprehensive analysis Of the
experiment’s outcomes .
Applications of Genetic Engineering:

- It has wide range of applications and their product is useful for Science, Medicine,
Agriculture and industry here are some application of Genetic Engineering.

Agriculture Revolution:

Genetic Engineering has led to create a genetically modified crops with enhanced resistance
to diseases and also improved nutritional content and increased yields.

2. Medicine Advancement:

Genetic engineering play vital role in developing novel medical treatments. From gene
therapy to personalized medicine, this field holds the promise of curing genetic disorders
and enhancing overall healthcare.

3. Environmental conservation:

Genetic engineering also helps in environmental conservation by facilitating the

development of organisms that can thrive in harsh conditions, helping to clean up polluted
environments, and also contribute to the preservation of endangered species.

4. Industrial Biotechnology: Genetic engineering also plays a vital role in industrial

applications, microorganisms are employed for efficient and sustainable manufacturing
processes and enabling the production of biofuels, pharmaceuticals, chemicals and other
valuable industrial products.

5. Bioremediation: Genetic engineering contributes to environmental cleanup through

bioremediation. Engineered microorganisms are designed to break down pollutants,
offering sustainable solutions for remediation of contaminated sites.

Refences:

www.Google.com

Botany C by Prof.Iftkhar Ahmed Khan & Prof.Riaz-ul-Haq Ramay