HSC Biology 1st Paper (Botany) Dr. Md.

Abdul Alim Sir

Book Page : 344-347
Chapter 11 (Biotechnology)
Akkarpatra : Page 397-400

Genetic Engineering:
The process by which an undesired part of a DNA molecules is excised off or a foreign
desired gene is added to the DNA molecule is known as Recombinant DNA technology or
Genetic Engineering.
In this process, the desired gene or the part of DNA is separated (by cutting at two desired
place) and joined to the specific part of another DNA molecule of the desired organism to
create a new gene combination. The technology involves in genetic engineering is called gene
cloning / recombinant DNA technology / in vitro recombinant DNA technology. The
organisms with new genetic characteristics prepared by recombinant DNA technology are known
as transgenic organisms / GMO (Genetically Modified Organism) / GEO (Genetically
Engineered Organism) / LMO (Living modified organism).
The aim of genetic engineering is transforming of a new gene to produce developed organisms.

Materials used in Genetic Engineering :

1. Desired Gene :
Desired gene is the gene of specific organism which is used in genetic engineering to develop
better quality products / desired organism.

2. Complementary DNA :
Complementary DNA (cDNA) in DNA is made from mRNA (Messenger RNA). This make
use of the enzyme reverse transcriptase, which does the reverse of transcription, it synthesis
DNA from an RNA template.

3. Restriction Enzyme :
The enzyme used to cut the DNA molecules (A,T,G,C) at specific points are known as
restriction enzyme. One or more types of restriction enzymes are present in each bacterial cell.
These enzymes cut the viral DNA molecules entering bacterial cell into 4-6 sequences. For this
reasons, restriction enzyme is called the molecular scissor.

4. DNA Ligase enzyme :

This enzyme repairs broke DNA by joining two nucleotides in a DNA strand. It is commonly
used in genetic engineering to do the reverse of a restriction enzyme. i.e. to join together
complementary restriction fragments.
5. Plasmid :
A plasmid is a small, circular, double-stranded extra-nuclear DNA molecule that is distinct
from a cell’s chromosomal DNA. Plasmids naturally exist in bacterial cells. Plasmids used in
genetic engineering are called vectors.

Process of genetic Engineering :

1. Selection and Isolation of desired gene or DNA part :
The 1st step is preparing recombinant DNA is the selection and isolation of the desired
DNA part. For this, the selected cell is 1st dissolved by the lytic enzymes to bring out cellular
substances. The DNA thus separated is purified by chemical treatment.

2. Selection of Vector DNA :

Vector are those DNA molecules that can carry a foreign DNA fragments when inserted into it.
An ideal vector is used as plasmid vector.

3. Cutting the desired DNA segment with Restriction Enzyme (RE) :

Restriction enzyme are considered as ‘molecular scissor’ in genetic engineering and applying
to cut out the desired DNA segment at specific sequences.

4. Insertion of the desired DNA segment (foreign DNA) into vector :

The isolated segment of desired DNA is put into medium with ligase enzyme and vector DNA.
Thus a recombinant DNA is prepared with the desired DNA segment.

5. Transfer of recombinant DNA into host cell :

Generally, the recombinant DNA is transferred to a suitable host cell (Bacteria) for
multiplication before use. The host is generally selected as the species from which vector
plasmid was collected. E.coli are used as cloning host.

6. Expression of desired DNA in the host :

After multiplication of the transformed bacteria in culture medium, it is evaluated whether the
desired DNA part has been successfully inserted to the host DNA or not.

7. Transfer of the recombinant DNA to the desired organism and evaluation :

The recombinant DNA thus prepared successfully are transferred to the desired organism and
transgenic organism are produced. In this process, recombinant DNA is introduced in the egg or
embryo cell and transgenic organism (micro-organism, plants and animals).