Biotechnology Principles and Processes Class 12 notes

 Biotechnology: Principles and processes


Biotechnology Principles and Processes Class 12 notes:




What is biotechnology?

 ● Biotechnology alludes to the innovation utilizing science, which has applications in horticulture, food handling industry, medication diagnostics, bioremediation, squander treatment, and energy creation.

● The European Federation of Biotechnology (EFB) characterizes biotechnology as "the joining of inherent science and creatures, cells, parts thereof and atomic analogs for items and administrations".

Basis of Modern Biotechnology

Genetic engineering − Introduction of unfamiliar hereditary material (DNA/RNA) into the host's genome and modifying its aggregate

Aseptic techniques − Involves support of contamination-free feeling in substance designing cycles for assembling of items like anti-infection agents, antibodies, etc. This is done as such as to empower the development of just wanted organisms answerable for a bioprocess.


Biotechnology Principles and processes Class 12 notes 

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Genetic Engineering

● Asexual reproduction saves the hereditary data while sexual multiplication jam varieties.

● Plant and creature hybridization methodology regularly bring about presentation of unfortunate qualities alongside advantageous ones.

● Genetic engineering conquers this constraint.

● Genetic engineering incorporates:

1.    Creation of recombinant DNA

2.    Gene cloning

3.    Gene transfer into host creature

 ● The presented piece of DNA doesn't repeat in the host except if it is coordinated with the chromosome of host.

● For getting replicated, the unfamiliar DNA should coordinate into the host DNA arrangement having 'beginning of replication'. At the point when this joining happens, unfamiliar DNA is repeated and many duplicates are shaped. This cycle is called cloning (the course of arrangement of different indistinguishable duplicates of DNA).

Biotechnology Principles and Processes Class 12 notes:

Development of a Recombinant DNA

 ● Plasmid (independently duplicating, roundabout, extra chromosomal DNA) is segregated.

● Plasmid DNA acts as a vector since it is utilized to move the piece of DNA appended to it to the host.

 ● Plasmid DNA additionally contains qualities answerable for giving anti-infection protection from the microscopic organisms.

● Plasmid DNA was cut with a particular restriction catalyst ('atomic scissors' − that cut a DNA at specific locations).

 ● The DNA of interest (to be embedded) was likewise cut with a similar restriction compound.

● The DNA of interest is hybridized with the plasmid with the assistance of DNA ligase to frame a Recombinant DNA.

 ● Recombinant DNA is then moved to a host like E.coli, where it imitates by utilizing the host's duplicating hardware.

● When E.coli is refined in a medium containing anti-toxin, just cells containing recombinant DNA will actually want to make due because of anti-toxin obstruction qualities and one will actually want to disconnect the recombinants.


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Restriction Enzymes as Tools of RDT

● Restriction proteins are specific chemicals that perceive and cut a specific succession of DNA.

● Nucleases are of two kinds:

o   Endonucleases − Cut the DNA at explicit situations inside the DNA

o   Exonucleases − Cut the DNA at the closures (Remove the nucleotides at the finishes of the DNA)

● Every restriction enzyme recognizes various arrangements (Recognition successions). More than 900 restriction proteins have been segregated, all of which perceive various successions.

● Recognition groupings are palindromic - Palindromes are the succession of base combines that read same both backward and forwards  (i.e., same and bearing).

● Restriction compounds remove a little from the focal point of palindrome site, however between similar two bases on the contrary strands.




● Thus, overhangs (called sticky ends) are produced on each strand.

● Sticky closures structure hydrogen bonds with their integral partners with assistance of DNA ligases.

● All these cycles structure the premise of RDT.

● Naming restriction enzyme

o   I st letter − Genus of the life form from which the enzyme is derived

o   II nd and III rd letters − Species of the organism

o   IV th letter − Name of the strain

o   Roman number − Order of seclusion E.g., In EcoRI − Derived from E.coli, strain R. It is the I st to be found.




Gel Electrophoresis

● The sections acquired in the wake of cutting with restriction compounds are isolated by utilizing gel electrophoresis.

● Electric field is applied to the electrophoresis grid (normally agarose gel) and adversely charged DNA parts move towards the anode.

 ● Fragments separate as per their size by the sieving properties of agarose gel. More modest the section, farther it moves.

● Staining colors, for example, ethidium bromide followed by openness to UV radiations are utilized to picture the DNA parts.

● DNA sections are noticeable as dazzling orange shaded groups in the agarose grid.

 ● These bands are cut from the agarose gel and extricated from the gel piece (elution).

● DNA sections are refined and these sanitized DNA pieces are utilized in building recombinant DNAs.




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Cloning vectors and host as apparatuses of RDT

Cloning Vectors

● Plasmids and bacteriophages are usually utilized as cloning vectors.

● Both of these can recreate inside the bacterial cells free of the chromosomal DNA.

Bacteriophages − Have high duplicate number (of genome) inside the bacterial cell

Plasmids − May have 1 − 2 duplicate number to 15 − 100 duplicate number for every cell

● If unfamiliar DNA is connected to these vectors, then, at that point, it is increased to the number equivalent to the duplicate number of vector.

● Features present in the actual vector help in the simple separation of recombinants from the non-recombinants.

Parts of a plasmid cloning vector

● Origin of replication (ori)

○ Replication begins from ori. Any part of DNA when connected to ori can be made to recreate.

○ With the assistance of this, the genetic engineer might control duplicate number of the recombinant DNA. To recuperate a big number, appropriate beginning of replication should be picked.

 ● Selectable marker

○ These qualities help to choose recombinants over non-recombinants.

○ Antibiotic opposition qualities like amp R (ampicillin resistant), tet R (tetracycline resistant) fill in as selectable markers normally.

Biotechnology Principles and Processes Class 12 notes:

Cloning sites

○ These locales allude to the acknowledgment locales for restriction compounds (like EcoRI, Hind III, PvuI , BamHI, and so on)

○ These are the sites where restriction catalysts cut the DNA.

○ Cloning process becomes finished when more than one acknowledgment sites are available.

○ Therefore, ligation is done distinctly at the restriction sites present on the antibiotic resistance genes.

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How anti-infection opposition qualities help in choosing recombinants?

● Suppose tet R quality has Bam HI acknowledgment site.

● When BamHI is utilized for restriction, foreign DNA section is embedded inside the tet R gene.

● Hence,  tetracycline. obstruction is absent in the recombinants.

● Recombinants will develop on the media containing ampicillin, yet will kick the bucket on media containing   tetracycline..

● On the other hand, non-recombinants will develop on medium containing ampicillin just as on medium containing   tetracycline.

 ● In this way, antibiotic resistance gene helps  in choosing transformants.

Substitute selectable marker

● Other than anti-infection obstruction qualities, elective markers can be utilized.

● One of them is quality coding for a^- galactosidase.

● When unfamiliar quality is embedded inside a^galactosidase quality, the catalyst a^¬galactosidase gets inactivated (insertional inactivation)

Then the microscopic organisms are become on a chromogenic substrate.

 ● Non-recombinants will deliver blue-colored provinces.

● Recombinants will deliver dry settlements.


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Cloning vectors for plants and creatures

● Ti plasmid (tumor-inducing plasmid) alludes to the plasmid of Agrobacterium tumefaciens.

o   A. tumefaciens is a plant microorganism. It produces growths in the plants it taints.

o   Ti plasmid can be adjusted into a cloning vector by eliminating the qualities liable for pathogenicity.

  Retrovirus − These are the infections that contaminate creatures. They produce malignant growths in creatures.

o   Retroviruses can be incapacitated to be utilized as a cloning vector.

competent host

● Competent host alludes to the bacterial cells that can take up the vector (containing Recombinant DNA).

 ● Methods to bring recombinant DNA into capable host:

o   Cells are treated with divalent cations (for example Ca 2+ ). Then, at that point, these cells are hatched with recombinant DNA on ice, trailed by heat shock (at 42º), and afterward setting them back aside momentarily. By this, microbes can take up recombinant DNA.

 Microinjection− Recombinant DNA is straightforwardly infused into the core of creature cell.

Biolistics (Gene Gun)− Cells are assaulted with high speed miniature particles of gold or tungsten.

 ○ Disarmed vector as if there should be an occurrence of A. tumefaciens and retrovirus.

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Biotechnology Principles and Processes Class 12 notes:


Processes of RDT

Isolation of Genetic Material (DNA)

● For the cycles of RDT, DNA should be accessible in its unadulterated structure.

● First of all, cells are treated with explicit synthetic compounds to tear open the cell to deliver cell parts like DNA, RNA, proteins, and so on This is finished by chemicals like lysozymes (bacterial cell), cellulase (plant cell), and chitinase (parasitic cell).

● Contaminants, for example, RNA and proteins are processed with the assistance of ribonucleases and proteases separately.

● Addition of chilled ethanol eventually hastens out the refined DNA, which can be viewed as assortment of fine strings in the suspension.

 Cutting of DNA at Specific Locations

● DNA is cut into pieces with the assistance of restriction proteins.

● Fragments produced after restriction are detached with the assistance of gel electrophoresis.

 ● Recombinant DNA is acquired by hybridizing 'quality of interest' with vector, with the assistance of enzyme DNA ligase.

Polymerase Chain Reaction (PCR)

● Recombinant DNA can be enhanced by PCR. A few indistinguishable duplicates of it tends to be combined in vitro.

● Two arrangements of primers (chemically combined oligonucleotide extends that are integral to a locale of DNA), enzyme DNA polymerase, and deoxynucleotides are added.

● PCR comprises of 3 stages:

Denaturation− Double helical DNA is denatured by giving high temperature. DNA polymerase doesn't get corrupted in such high temperatures since the DNA polymerase utilized in this response is thermostable as it is disconnected from thermophilic microscopic organisms, Thermus aquatics.

Extension − Replication of DNA happens in vitro.

This cycle is rehashed a few times to produce up to 1 billion indistinguishable duplicates of the DNA.

Addition of Recombinant DNA into Competent Cells

● Insertion of recombinant DNA into host is done by a few techniques:

 ○ Transformation if there should be an occurrence of microbes

○ Disarmed vectors, biolistics, and micro¬injections in the event of plant and creature cells

 ○ The cells bearing recombinant DNA are chosen on the grounds that the recombinants only have selectable marker present in them (like anti-toxin obstruction).

Acquiring the Foreign Gene Product

● This is the stage for which the recombinant DNA was created.

 ● The cell containing recombinant DNA will deliver a clever protein item (positive item/Recombinant protein).

● For enormous scope creation of the advantageous item (anti-toxins, immunizations, chemicals), ideal conditions are to be given.

● Continuous culture − Used culture media is depleted from one side and new culture media is added from the opposite side.

o   Cells are kept all through in their log/outstanding stage.

o   Larger biomass is delivered by this strategy prompting better return.

 ● Bioreactors − Large vessels in which enormous volumes (100 − 1000 liters) of culture can be created

 ○ Optimal development conditions for microorganisms are available (temperature, pH, substrate, salts, vitamins nutrients, and so on)

 ○ A bioreactor has the accompanying parts ¬ fomenter framework, oxygen conveyance framework, froth control framework, temperature and pH control framework, testing ports.


Biotechnology Principles and Processes Class 12 notes:


Downstream Processing

● Biosynthesis of many mixtures like proteins, alcohols, and anti-microbials occur inside the bioreactor.

 ● The items so acquired are unrefined and require partition, cleaning, and completing, which is done under downstream handling (DSP).

 ● DSP makes an unrefined bio item attractive.

 ● After legitimate division and purging, additives are added and the completed item is made to go through clinical preliminaries and quality checks prior to being shipped off market.

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