Genetic engineering Flashcards
is known to be the best known application of molecular genetics
Genetic engineering
Requirements for Recombinant DNA Technology (6)
- Restriction Endonuclease and DNA Ligase
- DNA ligase
- Vehicles for cloning
- Functional host cell
- A method for selection of recipient cells and their multiplication
- Recipient cells
part of cell’s defense mechanisms against invading foreign genomes
- Restriction Endonuclease and DNA Ligase
cut within certain well-defined based sequences in the invading DNA fragments
- Restriction Endonuclease and DNA Ligase
shown a high degree of specificity - possible to direct the splitting of DNA of the cloning vehicle
- Restriction Endonuclease and DNA Ligase
Enzyme that makes possible the insertion of selected piece of DNA
- DNA ligase
Commonly used vector/vehicles for molecular cloning: (3)
a. Plasmids
b. Bacteriophages
c. Agrobacterium tumefaciens
cells that show the characteristics of the introduced gene
- Recipient cells
refers to the joining together of DNA molecules from two different species that are inserted into a host organism to produce new genetic combinations that are of value to science, medicine, agriculture, and industry.
Recombinant DNA technology
is the general name for a piece of DNA that has been created by the combination of at least two strands.
Recombinant DNA (rDNA),
Steps of Genetic Recombination Technology (6)
- Isolation of Genetic Material
- Restriction Enzyme Digestion
- Amplification Using PCR
- Ligation of DNA Molecules
- Insertion of Recombinant DNA into the Host
- Isolation of Recombinant Cells
is a method of making multiple copies of a DNA sequence using the enzyme – DNA polymerase in vitro.
PCR
helps to amplify a single copy or a few copies of DNA into thousands to millions of copies.
PCR
PCR reactions are run on ‘thermal cyclers’ using the following components: (4)
Template
primers
enzyme
nucleotides
– DNA to be amplified
Template
small, chemically synthesized oligonucleotides that are complementary to a region of the DNA.
b) Primers
needed to extend the primers by the enzyme.
d) Nucleotides
The process of joining these two pieces together using the enzyme ‘DNA ligase’ is ‘
ligation
The resulting DNA molecule is a ___ of two DNA molecules – the __ and the __.
hybrid; interest molecule, vector
this new hybrid DNA molecule is also called a ____ and the technology is referred to as the _____
recombinant DNA molecule; recombinant DNA technology.
In this step, the recombinant DNA is introduced into a recipient host cell mostly, a bacterial cell. This process is ‘____’.
Transformation
For isolation of recombinant cell from non-recombinant cell, ___ of plasmid vector is employed.
marker gene
Researches on Human Genome (4)
-Mapping of Human Genome
-DNA Fingerprinting
-Identification, DNA Sequencing and determination of gene action of human genes
-Mitochondrial DNA and Aging
What year??
DNA sequencing together with computer software that align DNA pieces facilitated the
shift from gene mapping to DNA sequencing.
1995
What year ??
Data on human genome sequencing is shown in the public.
1996
What year??
Sequencing of 99% of the human genome accomplished.
2000
What year??
Progress in microarray technology (for investigating gene function and interactions)
2000 to present
technique of DNA fingerprinting involves the use of: (2)
-Restriction Fragment Length Polymorphisms (RFLPs),
-Variable Number of Tandem Repeats (VNTRs) or Minisatellites
regions of DNA that vary among individuals
Restriction Fragment Length Polymorphisms (RFLPs),
another method of DNA fingerprinting consider tandemly repeated sequences.
Variable Number of Tandem Repeats (VNTRs) or Minisatellites
The focus is on genes
that control heritable diseases such as: (4)
a) Osteoarthritis
b) Duchenne muscular dystrophy
c) Huntington’s disease
d) breast cancer disposition gene (BRCA 1 and BRCA 2)
It is claimed that aging may be caused by the buildup mutation of ____
mitochondrial DNA.
Researches on Animal Genes (4)
Super-ovulation Hormone
Rat Pancreatic Peptide Gene
Transgenic Fishes
Transgenic Livestock and Poultry
A recombinant ______ produced by Integrated
Genetics, Inc. causes superovulation in cattle.
bovine follicle-stimulating hormone (bFSH)
Injection of ____ into a strain of obese rats reduced feeding and absorption of fats
pancreatic peptide
This transgenic fish Carries foreign gene for growth hormone; grow faster than non-transgenics
salmon
In salmon, Alongside the growth hormone gene, the gene for ___ was inserted
beta galactosidase
To improve the characteristics of livestock and poultry, _____ are
transferred to the animals
growth hormones
30% more efficient in feed conversion Reach the marketable size 7 week earlier than their non-transgenic counterpart
Transgenic pigs
. Researches on Plant Genes (5)
Genetically Modified for resistance to insect pest, disease & herbicides
-Genetically modified for improved nutritional qualities
-Genetically modified for special products
-Genetically modified for enhanced quality traits
-Mapping of Plant Genomes
Insertion of _____ for crystals toxic to Lepidopteran insects
into canola, corn, cotton, potato, soybeans and tomato provide season-long protection against Lepidopteran insect pests.
Bacillus thuringiensis (Bt) gene
Insertion of _____ genes provides resistance against papaya ring spot virus
and banana bunchy top virus.
Virus coat protein
bar gene: _____
bxn genes: from ____
for resistance against ___.
Streptomyces hygrospicus
Klebsiella
herbicides
this gene is introduced into rice
Methionine-rich gene
Genes for enzyme that converts sucrose to ____ are inserted into sugar beets; yields __ of their dry weight as fructans.
fructans; 40%
genes from ___ flower and bacterium ____, the source of ____ enzyme to enable rice to synthesize ____, precursor of vitamin A
daffodil;
Erwinia uredovora;
phytoene desaturase;
carotene
iron- storage protein; accumulates in the endosperm, tripling the rice iron content.
Ferritin
Transgenic potatoes producing vaccine against (3)
E.coli enterotoxin, cholera toxin and Norwalk virus.
Transgenic bananas and tomatoes producing vaccines against ___
hepatitis B.
Transgenic tomatoes producing vaccines against respiratory ____ (causes lower respiratory tract disease in infant and children)
syncytial virus
Delayed tomato ripening by inserting antisense gene for ____ (enzymes for breaking down pectin)
polygalacturonase
tomatoes with ___ gene ripen on the vine to develop flavor and color before the softening process, thus prolonging its shelf life
antisense
Improved flavor and sweetness by inserting genes for ___ into tomato and lettuce.
moneliin
insertion of antisense gene for ____ (enzyme that polymerizes phenolic compound to form dark pigment)
polyphenol oxidase
Genome mapping requires identification of ___
DNA markers
– DNA fragments that can distinguished from one another due to their differences in nucleotide sequences.
DNA markers
Researches on Microbial Gene (4)
-Iron-oxidizing bacteria
-Microbes to track organisms
-Bacterial biosensors
-Molecular pharming
Iron-oxidizing bacteria is used to leach metals (Cu, uranium) from __
Fe ore
Used to process waste H2S gas produced by petroleum refineries
Iron-oxidizing bacteria
Bacteria genetically engineered to glow in the dark Can be used as living biosensors for detection of virtually any class of toxic agents
Bacterial biosensors
Hormone for glucose uptake; For diabetis
Insulin
Dissolves blood clot; heart attack victims
Tissue plasminogen activator (TPA)
Blood clotting factor; certain types of hemophilia patients
Factor VIII
Lower blood pressure; hypertension
Renin inhibitor
production of medically important proteins in the mammary gland of livestock
Molecular pharming
Dissolves blood clot
Host:__
Tissue plasminogen activator (TPA); Goat
Used as iron supplement in infant formula
Host:
Lactoferrin
Cattle
Treatment of diabetis
Host:
Insulin-like growth factor 1
Cattle
Biosafety and Ecological Implications (2)
- Potential ecological effects
- Regulatory policies
must be the basis for planning for the development, testing and risk assessment of the introduction of GE organisms into environment.
Ecological principles
