Flashcards

for NEET Biology: Biotechnology Principles and Processes

Manipulation and alteration of

the genetic makeup of
organisms using biotechnology

Genetic engineering Recombinant DNA (rDNA) is

used

An organism with a desired trait

is generated

Use of biotechnology in
chemical engineering processes
to manufacture products from
Bioprocess living organisms in a large
quantity
engineering
E.g. vitamins, vaccines,
antibiotics, etc.

Extrachromosomal, circular
DNA, which replicates
autonomously
Plasmid
E.g. commonly present in
bacteria
 Flashcards for NEET Biology: Biotechnology Principles and Processes

DNA formed by combining DNA

pieces or desired genes of two
different species or organisms
Recombinant DNA Stanley Cohen and Herbert
(rDNA) Boyer produced the first artificial
rDNA by adding antibiotic
resistant genes to the plasmid
of Salmonella typhimurium
Molecular scissors, restriction
endonucleases

They cut the DNA at a specific

palindromic sequence forming
sticky ends
Restriction enzymes
Werner Arber, Hamilton Smith,
and Daniel Nathans discovered

Hind II- the first restriction

endonuclease
Isolated from E.coli RY13

Recognition site, where the

enzyme cuts is G/AATTC
EcoRI (5’→3’) and in the
complementary strand it is
CTTAA/G (3’→5’)

Discovered in Boyer’s UCSF lab

 Flashcards for NEET Biology: Biotechnology Principles and Processes

Joins the sticky ends of DNA

fragments cut by restriction
enzymes
DNA Ligase
Joins Okazaki fragments of
lagging strand during replication

Father of genetic engineering

Carried out gene-splicing

experiments
Paul Berg
Added genes of lambda phage
virus into DNA of SV-40 virus

Received Nobel prize in 1980

DNA carriers for transferring the

desired gene or DNA fragment
into the host
Vectors
E.g. plasmids (pBR322, Ti),
bacteriophages (λ phage)
 Flashcards for NEET Biology: Biotechnology Principles and Processes

E.coli plasmid

Origin of replication- ori

Selectable markers- ampicillin and

pBR322 tetracycline resistant genes, i.e.
ampR and tetR

Restriction sites- BamH I, Sal I (in

tetR gene), Hind III, EcoR I, Pvu II,
Pst I (in ampR gene), Cla I, etc.
In vitro synthesis of multiple
copies of genes of interest,
invented by Kary Mullis

Polymerase chain The oligonucleotide primer is used

after denaturation- Annealing
reaction (PCR)
Primer is extended using
thermostable DNA polymerase,
i.e. Taq polymerase isolated from
Thermus aquaticus (bacterium)
Introducing foreign DNA into host
cell

Using vector

Without vector- microinjection (direct

Gene transfer injection of rDNA into the nucleus of
animal cells)

Biolistics or gene gun- plant cells are

bombarded with microparticles of
gold or tungsten coated with DNA
 Flashcards for NEET Biology: Biotechnology Principles and Processes

The process of cutting and

Elution extracting separated DNA
pieces from gel electrophoresis

Downstream The process of separation and

purification to make the finished
processing product after biosynthesis