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TRANSGENESIS - Coggle Diagram
TRANSGENESIS
Process of transferring FG into loving organism
Exhibit new property & transmit to offspring
Three type of genetic material
Double standard gene segment, oligonucleotides, cDNA sequences
Physical technique
Microinjection
use of micropipette to inject liquid into living cell and ICS
use inverted microscope with 200x
dissecting stereo microscope at 40-50x
method 2 transfer gene into animal
inject rDNA into pronuclei in embryo, 12-14 after fertilization, transfer the embryo into mother
method 1 inner cell
DNA is extract from embryonic stem cell, select cell for expression, inject into inner cell mass (blastocyst), implant in uterus
Biolistic
Microparticle gun use to accelerate microprojectiles deliver DNA into cell
DNA coated into microscopic bead coated w gold
coated bead attach to end of plastic bullet, loaded into firing chamber of gene gun
explosive force fire the bullet w bead toward target cell lie beyond of barrel
Electroporation
DNA delivered direct into cell w electrical pulse
EF cause the increasing permeability of cell membrane to allow chemical, drug or DNA to be introduced to the cell
Chemical technique
Lipid and liposome mediated
mix lipid w DNA, add to cell, (DNA encapsulated inside liposomes, lipo adhere to cell, lipo bilayer fuse w plasma membrane, DNA enter cell, DNA in nucleus), FG express
Calcium phosphate mediated
isolate DNA mix w cal chl & pota phos, incubated into cell, fraction of cell take the cal phos precipitate via endocytosis
Biological technique
Conjugation
natural transfection (survival of bacterium)(horizontal gene transfer)
cell w F plasmid can produce sex pilus and connected w recipient cell
transfer the gene to the recipient cell and the cell become new donor
Transformation
direct uptake of exogenous DNA through cell membrane by competent cell (ready to bring foreign DNA)
w DNA fragment
bacterial chromosome, take up the DNA, integration by nonreciprocal recombination, produce stable & degradation of enzyme form unstable transformation
w DNA plasmid
produce stable transformation
Retroviral transduction
DNA transferred from one bacterium to other by virus (viral vector)
lytic and lysogenic cycle
Agrobacterium mediated transfer
soil borne gram negative bacterium
invade dicot plants & cause crown gall disease
w presence of Ti plasmid (transport new genes into plant cell)
infect plants, transfer Ti plasmid into host cell, T-DNA (segment) integrate, cause abnormal proliferation of plant cell
direct synthesis of organic acid, opines
Ti plasmid infection
remove T-DNA and insert DNA w desired gene into Ti plasmid, transfer back to the agrobacterium and introduce to the plant