Biotechnology (principles and processes)

Question 1

Genetic engineering

Answer 1

Manipulation and alteration of the genetic makeup of organisms using biotechnology

Recombinant DNA (rDNA) is used

An organism with a desired trait is generated

Question 2

Bioprocess engineering

Answer 2

Use of biotechnology in chemical engineering processes to manufacture products from living organisms in a large quantity

E.g. vitamins, vaccines, antibiotics, etc.

Question 3

Plasmid

Answer 3

Extrachromosomal, circular DNA, which replicates autonomously

commonly present in bacteria

Question 4

Vector

Answer 4

DNA carriers for transferring the desired gene or DNA fragment into the host

E.g. plasmids (pBR322, Ti), bacteriophages (λ phage)

Question 5

Recombinant DNA (rDNA)

Answer 5

DNA formed by combining DNA pieces or desired genes of two different species or organisms

Question 6

Stanley Cohen and Herbert Boyer

Answer 6

produced the first artificial rDNA by adding antibiotic-resistant genes to the plasmid of Salmonella Typhimurium in 1972

Question 7

Restriction Enzymes

Answer 7

Molecular scissors, restriction endonucleases, Chemical knives

They cut the DNA at a specific palindromic sequence forming sticky ends

Question 8

who discovered restriction enzymes?

_________– was the first restriction endonuclease

Answer 8

Werner Arber, Hamilton Smith, and Daniel Nathans

Hind II

Question 9

EcoRI

Answer 9

Isolated from E.coli RY13

Recognition site, where the enzyme cuts is G/AATTC (5’→3’) and in the complementary strand it is CTTAA/G (3’→5’)

Discovered in Boyer’s UCSF lab

Question 10

pBR322

Answer 10

E.coli plasmid

Origin of replication- ori

Selectable markers- ampicillin and 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.

Question 11

Paul Berg

Answer 11

Father of genetic engineering

Carried out gene-splicing experiments

Added genes of lambda phage virus into DNA of SV-40 virus

Received Nobel prize in 1980

Question 12

DNA ligase

Answer 12

Joins the sticky ends of DNA fragments cut by restriction enzymes

Joins Okazaki fragments of lagging strand during replication

Question 13

DNA polymerase

Answer 13

Help in DNA cloning

Question 14

Polymerase chain reaction (PCR)

Answer 14

In vitro synthesis of multiple copies of genes of interest, invented by Kary Mullis

The oligonucleotide primer is used after denaturation- Annealing

Primer is extended using thermostable DNA polymerase, i.e. Taq polymerase isolated from Thermus aquaticus (bacterium)

Question 15

Gene transfer

Answer 15

Introducing foreign DNA into host cell

Using vector

Without vector- microinjection (direct 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

Question 16

Elution

Answer 16

The process of cutting and extracting separated DNA pieces from gel electrophoresis

Question 17

Downstream processing

Answer 17

The process of separation and purification to make the finished product after biosynthesis