Gene Technology: Molecular Cloning, Applications and Techniques Report

This report provides a detailed overview of gene technology, encompassing key concepts such as enzymes involved in molecular cloning, techniques for transgene conformation, and the components of chromosomes and chromatin. It explores the processes of transcription and gene silencing machinery in eukaryotes, along with the components of genome editing and the application of molecular vaccines. The report further delves into the application of molecular techniques in forensic science and the concept of nanobiotechnology, including its applications and examples. Additionally, it contrasts conventional and modern genetic engineering, providing a comprehensive understanding of the field. The report is supported by relevant figures and references, offering a thorough exploration of the subject matter.

Gene technology

Table of Contents
Enzymes, components and techniques that are involved in molecular cloning..........................1
Key technologies with respect to conformation of transgene......................................................1
Components of chromosomes and chromatin..............................................................................1
Components and steps in transcription........................................................................................2
Gene silencing machinery in eukaryotes.....................................................................................2
Components of genome editing...................................................................................................3
Molecular vaccines......................................................................................................................3
Applying molecular techniques in forensic science....................................................................3
Concept of Nanobiotechnology, its applications and examples..................................................4
Difference between Conventional and modern genetic engineering...........................................4
REFERENCES................................................................................................................................5

Table of Figures
Figure 1: Placement of chromosomes and chromatin in eukaryotic cells.......................................2

Enzymes, components and techniques involved in molecular cloning
Enzymes involved in molecular cloning are:
 Type A# Alkaline Phosphatase
 Type B# Terminal Transferase
 Type C# Thermo-stable DNA Polymerases
 Type D# Bacteriophage RNA Polymerases
 Type E# Nucleases: DNase and RNase
 Type F# Polynucleotide Kinase
 Type G# DNA Ligase
 Molecular cloning is comprised of two main components. They are:
 DNA particle of interest which is to be replicated
 A vector backbone which has all other components of replication.
Techniques of molecular cloning are biolistics, optical injection, chemical sensitivity of cells
and electroporation (Waddington, 2014)
Key technologies with respect to conformation of transgene
Cre / LoxP is the common technology used to transgene. Some other technologies include
NGS, iPCR and Splinkerette for transgene mapping and targeted locus amplification (TLA)
which are used in crosslinking, fragmentation, relegation, etc.
Components of chromosomes and chromatin
Important components of eukaryotic chromosomes are DNA, RNA, metallic ions, histone
and non-histone proteins, protamines etc.
Chromatin is the loose yet dense network of chromosomes that are uncoiled and are
present in cell’s nucleus. It is only found in eukaryotic cells. The main component of Chromatin
is histones.
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Figure 1: Placement of chromosomes and chromatin in eukaryotic cells
(Source: Nucleic Acids, 2016)
Components and steps in transcription
Transcription is the process which is responsible for copies to mRNA. It helps in carrying
the information so as to conduct synthesis of protein.
There are three main events of transcription:
 Initiation: In this step, RNA polymerase and double stranded DNA are combines together. It
is an important step in the expression of gene.
 Elongation: At this stage, short stretch of DNA is developed which is actually single
stranded (Ohno, 2013).
 Termination: At this stage of transcription, RNA polymerase is released.
Gene silencing machinery in eukaryotes
Gene silencing machinery is used to silence the genes which reduces its expression. siRNA
and miRNA are short duplex RNA molecules which helps in exerting silencing effects. Both the
RNA’s are involved in epigenetics with the help of a process called, RNA-induced
transcriptional silencing or RITS. The difference between the two is that siRNA enters from
vectors, that is, viruses and miRNA does not. Further, siRNA is double stranded RNA while
miRNA is single stranded RNA.
Some of the common components used in it includes, RNAi, siRNA and CRISPR.
Argonaut is a vital component of RNAi pathway. The common applications of Gene silencing
machinery are:
 It is commonly used by researcher to understand the gene disorder among individuals.
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 Assessment of viral and bacterial genes
 Gene involvement in respiratory diseases (Schwartzentruber and et.al., 2012).
Components of genome editing
Genome editing is considered as one of the types of genetic engineering where DNA can
be inserted, deleted or replaced from the genome of living organism with the help of molecular
scissors. Common components of genome editing include the following:
 Targeted gene insertion: It is a homologues recombination in order to change endogenous
viral elements. The method helps in deletion of gene, removal of exons, addition of it and
intruding point mutation.
 Double strand breaks: It is the situation where both the strands of double helix are severely
damaged which can lead to gene rearrangements. There methods helps to repair the damages
strands are, non-homologous end joining (NHEJ), microhomology-mediated end joining
(MMEJ), and homologous recombination (Cruz, 2013).
 Homologues recombination: It is the process which requires presence of identical or nearly
similar sequence that can be used as a template to recover breakage in two strands.
Molecular vaccines
Vaccines help in prevention of the diseases and considered as one of the cost-effective
methods. It assists in providing genetically modified outputs. Further, It helps in preventing from
any antigens that can enter into the body. It further assists in protecting the genes from any
reordering or rearranging deformity that can affect its functioning.
Applying molecular techniques in forensic science
Forensic science helps in establishing the truth by identifying and comparing traces being
left at crime scene. It is the medico legal identification of criminalities. Molecular genetic
techniques in case of forensic identification can be applied in the following manner:
 Assessment of liquid or dried deposited blood
 Examination of various biological secretions such as saliva or semen
 Assessment of hard tissues, hair follicles, etc
It acts as a wide source to examine DNA of an individual.
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Concept of Nanobiotechnology, its applications and examples
Nanobiotechnology is the intersection of biology and nanotechnology. It helps in creating
and imagining the systems that can further be utilized for biological research. Some of the
common applications of nanobiotechnology includes:
 Performing clinical tests which help in revealing the presence of disease or any antibody
 Assessment of genetic composition of an individual
 Helps in identification of sparse cells which are rare
 Assists in developing drugs for the diseases which are conventional
 Plays an important role in the field of dentistry (MacArthur and et.al., 2012).
Difference between Conventional and modern genetic engineering
Genetic engineering is manipulating current genes which change make up of cells. It
includes transferring of genes, deletion and insertion of new genes as well. The conventional
genetic reengineering does not include technological equipment that is effective enough to
perform the functions. The chances to failure were higher. However, in case of modern genetic
engineering, chances to success is higher as all the modern equipment are used to perform
functions.
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REFERENCES
Books and Journals
Cruz, C. D., 2013. Genes: a software package for analysis in experimental statistics and
quantitative genetics. Acta Scientiarum. Agronomy. 35(3). pp.271-276.
MacArthur, D.G. and et.al., 2012. A systematic survey of loss-of-function variants in human
protein-coding genes. Science. 335(6070). pp.823-828.
Ohno, S., 2013. Sex chromosomes and sex-linked genes (Vol. 1). Springer Science & Business
Media.
Schwartzentruber, J. and et.al., 2012. Driver mutations in histone H3. 3 and chromatin
remodelling genes in paediatric glioblastoma. Nature. 482(7384). pp.226-231.
Waddington, C. H., 2014. The strategy of the genes (Vol. 20). Routledge.
Online
Nucleic Acids. 2016. [Online]. Available through <
https://courses.lumenlearning.com/boundless-biology/chapter/nucleic-acids/ >.
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