Assignment No.

BIO203: METHODS IN MOLECULAR BIOLOGY

SPRING 2022

Student Name: Fazilat Jaffri

Id: bc200413833

What are restriction enzymes? Also, explain the applications of restriction enzymes in
molecular biology.

Restriction Enzymes:

A restriction enzyme or restriction endonucleaseis a catalyst that cuts DNA into pieces at or
close to unambiguous acknowledgment locales inside particles known as restriction
sites.Restriction enzymes are one class of the more extensive endonuclease gathering of
enzymes. To cut DNA, all restriction enzymes make two entry points, when through each sugar-
phosphate spine (for example each strand) of the DNA twofold helix.

Recognition site:

Restriction enzymes perceive a particular succession of nucleotides and produce a twofold

abandoned cut in the DNA. The acknowledgment successions can likewise be characterized by
the quantity of bases in its recognition site, normally somewhere in the range of 4 and 8 bases,
and the quantity of bases in the grouping will decide how frequently the site will show up by
chance in some random genome, e.g., a 4-base pair arrangement would hypothetically happen
once every 4^4 or 256bp, 6 bases, 4^6 or 4,096bp, and 8 bases would be 4^8 or 65,536bp.Many
of them are palindromic, meaning the base grouping peruses something similar.

Types of Restriction Enzymes:

Restriction enzymes are traditionally classified into four types on the basis of subunit
composition, cleavage position, sequence specificity and cofactor requirements.

Type I Enzymes:

Type I enzymes are complex, multisubunit, blend restriction-and-modification enzymes that cut
DNA at arbitrary a long way from their recognition sequences.These enzymes require both ATP
and S-adenosyl-L-methionine to work; multifunctional protein with both restriction processing
and methylase activities.Type I enzymes are of considerable biochemical interest, yet they have
little practical value since they don't create discrete restriction parts or unmistakable gel-banding
designs.
Type II Enzymes:

Type II enzymes cut DNA at characterized positions near or inside their recognition groupings.
Most require magnesium; single capability enzymes free of methylase.They produce discrete
limitation sections and particular gel banding examples, and they are the overwhelming class
utilized in the lab for routine DNA examination and quality cloning. Instead of shaping a solitary
group of related proteins, Type II enzymes are a collection of inconsequential proteins of various
sorts. Type II enzymes much of the time contrast so totally in amino corrosive arrangement from
each other, and for sure from each and every other known protein, that they represent the class of
quickly advancing proteins that are in many cases demonstrative of association in have parasite
collaborations.

Type III Enzymes:

Type III enzymes are additionally huge blend limitation and-modification enzymes. They divide
beyond their recognition sequences and require two such groupings in inverse directions inside a
similar DNA particle to achieve cleavage; they seldom give total digests.Require ATP (yet don't
hydrolyse it); S-adenosyl-L-methionine invigorates the response however isn't needed; exist as a
component of a complex with a change methylase.

Type IV Enzymes:

Type IV enzymes perceive altered, normally methylated DNA and are exemplified by the
McrBC and Mrr frameworks of E. coli.

APPLICATIONS OF RESTRICTION ENZYMES

Restriction endonucleases are widely used in molecular biology research for the following
applications:

Genetic Engineering:

The most well known use of limitation endonucleases is as an instrument for genetic
engineering. The endonuclease action empowers control of the genome as well as presentation of
successions of interest in the host living being. This outcomes in the creation of the ideal quality
item by the host. This idea has extensive variety of utilizations in biotechnology in the creation
of anti-microbials, antibodies, enzymes, and a few optional metabolites.

DNA mapping:

DNA mapping utilizing restriction enzymes is a technique to get primary data of the DNA
piece. In this procedure the DNA is processed with a progression of restriction enzymes to
deliver DNA sections of different sizes. The resultant sections are isolated by agarose gel
electrophoresis and the distance between the restriction enzyme sites can be assessed. This can
be utilized to decide the design of an obscure DNA part.

Gene Sequencing:

A huge DNA particle is digested utilizing restriction enzymes and the subsequent sections are
handled through DNA sequencer to get the nucleotide sequence.