Biotechnology: Principles and Processes Flashcards
_______ deals with techniques of using live organism or enzymes from organism to produce products and processes useful to humans
Biotechnology
Techniques to alter chemistry of genetic material in order to change the phenotype to host organism are a part of _______ engineering
genetic
The two core techniques that enabled birth to modern biotechnology are :
Genetic and bioprocess engineering
Maintenance of sterile environment in chemical engineering processes to enable growth of desired organism in large quantities for manufacture of biotechnological products is called:
bioprocess engineering
The type of reproduction which permits variation is :
sexual reproduction
The type of reproduction which preserves genetic information is:
asexual reproducton
The techniques of genetic engineering which include creation of _______ use of ______ and ______, allows us to isolate and introduce only one or a set of desirable genes without introducing undesirable genes into the target organism
Recombinant DNA, gene cloning, gene transfer
The process of making multiple identical copies of any template DNA is called :
Cloning
The specific DNA sequence of chromosome responsible for initiating DNA replication is :
Origin of replication
First recombinant DNA was constructed in 1972 by ______ and ______
Stanley Cohen; Herbert Boyer
An autonomously replicating circular extra-chromosomal DNA of bacteria is called :
PLasmid
First recombinant DNA was constructed by linking a gene encoding antibiotic resistance with native plasmid of :
Salmonella typhimurium
Restriction enzymes are also known as :
Molecular scissors
The cutting of DNA at specific locations is done with the help of :
restriction enzymes
In genetic engineering, plasmid DNA act as ______ to transfer the piece of DNA linked to it into host organism
vector
The linking of antibiotic resistance gene with the plasmid vector became possible with the enzyme _____
DNA ligase
Replication of recombinant DNA in host is accomplished by using which enzymes of host?
DNA polymerase
Key tools of genetic engineering are:
Restriction enzymes, polymerase, ligase, vetor and host organism
The first restriction endonuclease that was found to cut DNA at specific sequence was-
Hind II
The specific base sequence recognised by restriction endonuclease to cut the DNA is called:
recognition sequence
Restriction enzyme Escherichia coli
RY 13 is written as:
EcoRI
While naming restriction enzymes, the order in which enzyme was isolated from the strain of bacteria is indicated by:
roman numbers
Restriction enzymes belong to which class of enzymes?
nucleases
Nucleases which remove nucleotide from the ends of DNA are called:
exonucleases
Nucleases which make cuts at specific position within DNA re called:
endonucleases
The sequence recognised by restriction endonuclease to cut the strand in DNA is:
palindromic nucleotide sequence
The sequence of base pairs that reads same on the two strands when orientation of reading is kept same is called :
Palindrome
The overhanging stretches or single stranded portion left at the end by restriction enzymes are called:
sticky ends
The stickiness of sticky ends facilitates the action of enzyme _____ on them
DNA ligase
Source DNA and ______ are need to be cut with same restriction enzyme, in order to create a recombinant molecule
vector
The fragments of DNA can be separated by a technique known as:
Gel electrophoresis
The charge on DNA molecules is :
negative
Most commonly used gel matrix for electrophoresis is:
agarose
Agarose is extracted from:
sea weeds
DNA fragments in agarose gel separate through sieving effect according to their:
size
During separation, agarose gel provides ______ effect
sieving
Compound used to visualise separated DNA fragments in agarose gel is:
ethidium bromide
Ethidium bromide is visible when gel is exposed to:
UV light
The process of cutting out and extracting separated bands of DNA from gel is called:
Elution
Bacteriophage have very ______ copy numbers of thier genome within bacterial cells
high
The sequence from where replication starts is called:
Origin of replication(ori)
Copy number of the linked DNA is under control of which sequence?
ori
_____ helps in identifying and eliminating non - transformants and selectively permitting the growth of the transformants?
selectable marker
The process through which a piece of DNA is introduced in a host bacterium is called:
Transformation
Genes encoding resistance to ____ are considered useful selectable markers
antibiotics
Name some antibiotic genes useful as selectable marker for E. coli.
Ampicillin, chloramphenicol, tetracycline, kanamycin
During cloning, the ligation of alien DNA is carried out at a restriction site present in ______ genes
antibiotic resistance
An enzyme used as a selectable marker for selection of recombinants from non - recombinants is:
Beta galactosidaes
During cloning, rDNA is inserted within coding sequence of beta - galactosidase, thus leading to:
insertional inactivation
While using insertional inactivation, the colonies which donot produce any colour in presence of chromogenic substrate are identified as:
Recombinants
Non-recombinants colonies appear ____ in presence of chromogenic substrate when subjected to insertional inactivation
blue
_______ of Agrobecterium is used as a modified cloning vector to deliver gene of interest into plants
Ti plasmid
The viruses which have capability to transform normal cells to cancerous cell in animals are
retroviruses
Agrobacterium delivers a piece of DNA known as _____ in plants to transform normal cells into tumor cells
T-DNA
A pathogen of dicot plants which is used as cloning vector is:
Agrobacterium tumifaciens
The cation used to make bacterial cells competent to take up DNA is
calcium
Heat shock is given to bacterial cells at which temperature to make them competent to take up DNA?
42 degree C
Recombinant DNA is directly injected into the nucleus of an animal cell by
micro-injection
In biolistic method of rDNA transfer, micro-particles of _______ are coated with DNA
tungsten or gold
Cells are bombarded with high velocity micro-particles of gold or tungsten coated with DNA in a method known as:
biolistics or gene gun
Genetic material of all organism is:
nuclei acid
To isolate DNA from bacteria enzymes used to break the cell open is :
Lysozyme
Isolation of DNA from plant cells is done by disintegrating the cell wall using enzyme:
cellulase
To isolate DNA from fungus, enzyme used to break the cell open is:
chitinase
Molecules of DNA are intertwined with proteins like:
histones
During isolation od DNA, DNA is made free from RNA by using enzymes:
Ribonuclease (RNase)
During isolation of DNA, DNA is made free from proteins by using enzymes:
Proteases
After isolating DNA, purified DNA is observed as fine threads in suspension when treated with chilled _______
ethanol
After restriction digestion, the technique which is employed to check whether the DNA has been digested is:
Agarose gel electrophoresis
Joining of source DNA and vector DNA is done with the help of enzymes:
DNA ligase
PCR stands for:
polymerase chain reaction
Multiple copies of the gene of interest are synthesised in vitro using two sets of primers in:
polymerase chain reaction
In PCR, the small chemically synthesised oligonucleotides that are complementary to the regions of DNA are called:
primers
DNA polymerase extends the _____ using nucleotides provided during PCR
primers
Thermostable DNA polymerase used in PCR is isolated from:
Thermus aquaticus
Ampicillin resistance gene when used to select transformed cell is called:
selectable marker
When a protein encoding gene is expressed in a heterologous host, it is called:
Recombinant protein
For production of recombinant protein, the type of culture system which produces a large biomass is:
continuous culture system
The vessels in which large volumes of raw materials are biologically converted into specific products under optimum conditions are called:
Bioreactors
The most commonly used bioreactors are:
stirring type
Stirrer in stirred-tank reactor facilitates even mixing and ______ availability
oxygen
The process of downstream processing involves _____ and ______
separation; purification
Identify A, B, C and D in the following figure
A - Vector DNA; B- EcoRI; C- Sticky end; D-Recombinant DNA
Mark A, B, C and D in the following figure
A- Vector DNA; B-Ligases; C-Recombinant DNA molecule;D-Transformation
Mark A and B in the following Figure
A-largest; B- Smallest
Identify the vector
pBR322
Mark A, B, C, D and E in the following figure
A-EcoRI; B-BamH I; C-ori; D-ampR; E-tetR
Mark A, B, C, D and E in the following figure
A-Denaturation, B-Primers, C-Annealing, D-Taq Polymerase+deoxynucleotides, E-extension
Mark A, B, C, D and E in the following figure
A-Motor, B-Foam Braker, C-Flat bladed impeller, D- Culture broth, E-Sterile air
Making curd, bread or wine, which are all microbe mediated processes, could also be thought of as a form of ______
biotechnology
In vitro fertilisation leading to a ‘test-tube’ baby, synthesising a gene and using it, developing a DNA ______ or correcting a _______ gene, are all part of biotechnolgy
vaccine, defective
What does EFB stands for?
European Federation of Biotechnology
The EFB has given a definition of biotechnology that encompasses both ____ view and modern _____ biotechnology
traditional, molecular
The definition given by EFB is as follows:- ‘ The integration of _____ and _____ cells, parts thereof, and molecular analogues for products and services’
natural science, organisms
_______ procedures used in plant and animal breeding, very often lead to inclusion and multiplication of undesirable genes along with desired genes
Traditional hybridisation
Foreign DNA, when it gets integrated into the _____ of the recipient, may multiply and be inherited along with the host DNA
genome
For the _____ of any alien piece of DNA in an organism, it is needed to be a part of a chromosome(s) which has a specific sequence known as ‘origin of replication’
multiplication
The construction of the first recombinant DNA emerged from the possibility of linking a gene encoding ______ with a native plasmid
antibiotic resistance
______ acts as an insect vector to transfer the malarial parasite into human body
mosquito
Escherichia coli, is a bacterium closely related to ______
Salmonella
The ability to multiply copies of antibiotic resistance gene in E. coli was called _____ of antibiotic resistance gene in E. coli
cloning
In the year 1963, the two enzymes responsible for restricting growth of _____ in Escherichia coli were isolated
Bacteriophage
Out of two enzymes discovered in 1963, one of these added ______ to cut DNA, while the other cut DNA
methyl groups
It was found that Hind II always cut DNA molecules at a particular point by recognising a specific sequence of _____ base pairs
six
Besides Hind II, today we know more than _____ restriction enzymes that have been isolated from over _____ strains of bacteria each of would recognise different recognition sequence
900, 230
The convention for naming these enzymes is the first letter of the name comes from the ______
genus
The second two letters while naming an enzyme comes from the species of the ______ from which they were isolated
prokaryotic cell
Each restriction endonuclease functions by ‘inspecting’ the _____ of a DNA sequence
length
Restriction endonuclease will bind to the DNA and cut each of the two strands of the double helix at specific points in their ______ backbones
sugar-phosphate
EcoRI cuts the DNA between bases G and A only when the sequence _____ is present in the DNA
GAATTC
Restriction enzymes cut the strand of DNA a little away from the ______ of the palindrome sites, but between the same two bases on the _____ strands
centre, opposite
Sticky ends are named so because they form ______ with their complementary cut counterparts
hydrogen bonds
______ are used in genetic engineering to form ‘recombinant’ molecules of DNA, which are composed of DNA from different sources/genomes
Restriction endonucleases
When cut by the same restriction enzyme, the resultant DNA fragments have the same kind of ________
‘sticky ends’
Ligases join foreign DNA to ______
plasmid
The cutting of DNA by restriction endonucleases results in the ____ of DNA
fragments
DNA can be separated by forcing them to move towards the _____ under an electric field through a medium/matrix
anode
You cannot see pure DNA fragments in the ______ light and without staining
visible
DNA appears as bright _____ coloured bands in a ethidium bromide stained gel exposed to UV light
orange
The DNA fragments purified in this way are used in constructing _____ by joining them with _______
recombinant DNA, cloning vectors
______ and _______ have the ability to replicate within bacterial cells
plasmids, bacteriophages
Some plasmids may have only one or two copies per cell whereas others may have _____ copies per cell
15-100
If we are able to link an alien piece of DNA with bacteriophage or plasmid DNA, we can multiply its number equal to the ______ of the plasmid or bacteriophage
copy number
Vectors used at present , are engineered in such a way that they help easy linking of _____ and ______ of recombinants from non-recombinants
foreign DNA, selection
If one wants to recover many copies of the target DNA it should be cloned in a vector whose origin support _____ copy number
high
In order to link the alien DNA, the vector needs to have very few, preferably _____ recognition sites for the commonly used restriction enzymes
single
Presence of more than one ______ within the vector will generate several fragments, which will complicate the gene cloning
recognition sites
You can ligate a foreign DNA at the ____ site of tetracycline resistance gene in the vector pBR322
BamH I
rop codes for the proteins involved in the ______ of the plasmid
replication
The recombinant plasmids will loose tetracycline resistance due to ______ of foreign DNA at BamH I site
insertion
In a vector, one antibiotic resistance gene helps in selecting the ____, whereas the other antibiotic resistance gene gets ‘inactivated due to insertion’ of alien DNA, and helps in selection of ______
transformants, recombinants
Selection of recombinants due to in activation of antibiotics is a cumbersome procedure because it requires _______ plating on two plates having _____ antibiotics
simultaneous, different
Alternative selectable markers have been developed which differentiate ______ from _______ on the basis of their ability to produce _______ in the presence of a chromogenic substrate
recombinants, non-recombinants, colour
We have learnt the lesson of transferring genes into plants and animals from______ and ______
bacteria, viruses
The tumor inducing (Ti) plasmid of Agrobacterium tumifaciens has noe been modified into a cloning vector which is no more ____ to the plants
pathogenic
Retroviruses have also been ______ and are now used ______ to deliver ______ genes into animal cells
disarmed, desirable
Once a gene or a DNA fragments has been ______ into a suitable vector it is transferred into a bacterial, plants or animal host (where it multiples)
ligated
Since DNA is a molecule, it cannot pass through cell membranes
hydrophilic
In order to force bacteria to take up the plasmid, the bacterial cells must first be made _______ to take up DNA
competent
Calcium increases the ______ with which DNA enters the bacterium through _____ in its cell wall
efficiency, pores
In heat shock recombinant DNA can be forced into cells by ______ the cells with recombinant DNA on ice, followed by placing the briefly at 42 C and then putting them back on ______
incubating, ice
Biolistics or gene gun method is suitable for _____
plants
________ vectors, which when allowed to infect the cell, transfer the recombinant DNA into the host
Disarmed pathogen
In majority of organisms the nuclei acid is ______ or DNA
deoxyribonucleic acid
In order to cut the DNA with restriction enzymes, it needs to be in pure form, free from other ______
macro-molecules
Since the DNA is enclosed within the ______, we have to break the cell open to release DNA
membranes
DNA is released along with other macromolecules such as RNA, proteins, _______ and also ______
polysaccharides, lipids
Genes are located on long molecules of DNA interwined with proteins such as ____
histones
DNA that separates out can be removed by ______
spooling
DNA ultimately precipitates out and can be seen as collection of ______ in the suspension
fine threads
Restriction enzymes digestions are performed by incubating _______ DNA molecules with the restriction enzymes, at the optimal conditions for that ______ enzymes
purified, specific
DNA is negatively charged molecule, hence it moves towards the ______ electrode
positive
The ______ of DNA involves several processes
joining
The cut out ‘______’ from the source DNA and the cut vector with space are mixed and ligase is added. This results in the preparation of ________
gene of interest, recombinant DNA
DNA polymerase uses _____ as template in PCR
genomic DNA
If the process of replication of DNA is repeated many times, the segment of DNA can be amplified to approximately ______ times
billion
Thermostable DNA polymerase remain active during the high temperature induced _______ of double stranded DNA
denaturation
The amplified fragment if desired can now be used to ligate with a vector for further ______
cloning
______ cells after making them ‘competent’ to recieve, take up DNA present in its surrounding
Recipient
When you insert a piece of alien DNA into a cloning vector and transfer it into a bacterial, plant or animal cell, the alien DNA gets _____
multiplied
In almost all recombinant technologies, the ultimate aim is to produce a desirable _______
protein
There is a need for the recombinant DNA to be ______
expressed
The ______ gene gets expressed under appropriate conditions
foreign
Expression of the target protein can be induced after cloning the ____ and having optimised the conditions
gene of interest
To induce the expression of the target protein, one has to consider producing it on a ______
large scale
The cells harbouring cloned genes of interest may be grown on a ______ in the laboratory
small scale
The cultures may be used for ______ the desired protein
extracting
The cultures can be purified it by using different ______ techniques
separation
In a continuous culture system wherein the _____ medium is drained out from one side while _____ medium is added from the other medium
used, fresh
The purpose of adding a fresh medium in continuous culture system is to maintain the cells in their physiologically most active ______ phase
log/exponential
______ cultures cannot yield appreciable quantities of products
small volume
What is the range of volume of culture that can be processed in a bioreactor?
100-1000 litres
A bioreactor provides the optimal conditions for achieving the ______ by providing optimum growth conditions
desired product
The optimum growth conditions in a bioreactor include optimum temperature, ______, substrate, ______, ______, oxygen
pH, salts, vitamins
A stirred-tank reactor is usually _____ or with a ______ base to facilitate the mixing of the reactor contents
cylindrical, curved
Alternatively air can be ______ through the reactor
bubbled
Bioreactor has an ______ system, an ______ delivery system and a ______ control system, a ______ control system, ____ control system and sampling ports
agitator, oxygen, foam, temperature, pH
A bioreactor has sampling ports so that small volumes of the culture can be _____ periodically
withdrawn
After completion of the _____ stage, the product has to be subjected through a series of processes before it is ready for marketing as a ____
biosynthetic, finished product
The product has to be formulated with suitable ______
preservatives
Formulation has to undergo through clinical trails as in case of ____
drugs
Strict ______ testing for each product is also required
quality control
The ______ and _____ testing vary from product to product
downstream processing, quality control
The purpose of steam in a simple stirred-tank bioreactor is ______
sterilisation
Surface area is increased in a bioreactor for ____ transfer
oxygen
_____ dramatically increase the oxygen transfer area
Bubbles
What does the following figure depicit?
Removal of separated DNA by spooling
