Genetic Engineering
GENETIC ENGINEERING
GENETIC ENGINEERING GENERAL STEPS
What is Genetic Engineering?
Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a particular protein.
Ex. Inserting the human insulin gene into bacteria to mass produce it.
General Steps
Obtain the gene of interest (ex. insulin gene)
Insert the gene into the host (ex. bacteria)
Allow the host to multiply and express the foreign gene get your desired protein!
Get lots of cells that can make the protein = clones
The Big Picture
The inserted gene is transcribed and translated using the RNA Polymerase, ribosomes and other resources in the cell
Plasmids
Circular DNA
Extrachromosomal
NOT part of the E. coli genome
“extra” DNA
Contain a few non-essential genes
Can give the bacteria additional “traits”
Depends on the genes on the plasmid
Can be exchanged between bacteria
Recombinant plasmids
Plasmids can be modified in biological labs
Modified plasmid = Recombinant plasmid
Plasmids can be used as cloning vectors to get the recombinant plasmid into E. coli
Cloning vectors = way to get the gene of interest into the host
Transformation
Process in which foreign DNA is physically inserted into host E. coli cells.
E. coli that contains recombinant plasmid = Transformed cell
Transformation Steps
Recombinant plasmids and host E. coli are mixed together
CaCl2 is added
The Ca2+ ions neutralize the negative charges on plasmid DNA
Help plasmid enter the membrane
Transformation Steps
Heat Shock
By rapidly changing the temperature of the solution, temporary pores are opened in the membrane
Creates an opening for the plasmids to enter the E. coli
Transformation
Transformation is not 100% successful
After transformation
Some cells will contain plasmid = transformed
Some cells won’t contain plasmid = untransformed
In a later step, you will determine which cells were transformed
E. Coli as a host
E. coli is a good host because:
GENETIC ENGINEERING GENERAL STEPS
What is Genetic Engineering?
Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a particular protein.
Ex. Inserting the human insulin gene into bacteria to mass produce it.
General Steps
Obtain the gene of interest (ex. insulin gene)
Insert the gene into the host (ex. bacteria)
Allow the host to multiply and express the foreign gene get your desired protein!
Get lots of cells that can make the protein = clones
The Big Picture
The inserted gene is transcribed and translated using the RNA Polymerase, ribosomes and other resources in the cell
Plasmids
Circular DNA
Extrachromosomal
NOT part of the E. coli genome
“extra” DNA
Contain a few non-essential genes
Can give the bacteria additional “traits”
Depends on the genes on the plasmid
Can be exchanged between bacteria
Recombinant plasmids
Plasmids can be modified in biological labs
Modified plasmid = Recombinant plasmid
Plasmids can be used as cloning vectors to get the recombinant plasmid into E. coli
Cloning vectors = way to get the gene of interest into the host
Transformation
Process in which foreign DNA is physically inserted into host E. coli cells.
E. coli that contains recombinant plasmid = Transformed cell
Transformation Steps
Recombinant plasmids and host E. coli are mixed together
CaCl2 is added
The Ca2+ ions neutralize the negative charges on plasmid DNA
Help plasmid enter the membrane
Transformation Steps
Heat Shock
By rapidly changing the temperature of the solution, temporary pores are opened in the membrane
Creates an opening for the plasmids to enter the E. coli
Transformation
Transformation is not 100% successful
After transformation
Some cells will contain plasmid = transformed
Some cells won’t contain plasmid = untransformed
In a later step, you will determine which cells were transformed
E. Coli as a host
E. coli is a good host because: