Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of...
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Transcript of Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of...
![Page 1: Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a.](https://reader036.fdocuments.in/reader036/viewer/2022072011/56649e0c5503460f94af55d4/html5/thumbnails/1.jpg)
Genetic Genetic EngineeringEngineering
![Page 2: Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a.](https://reader036.fdocuments.in/reader036/viewer/2022072011/56649e0c5503460f94af55d4/html5/thumbnails/2.jpg)
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.
Image taken without permission from http://www.medicalprogress.org/uploads/images/insulin%20inject%20WSU%20210.jpg
![Page 3: Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a.](https://reader036.fdocuments.in/reader036/viewer/2022072011/56649e0c5503460f94af55d4/html5/thumbnails/3.jpg)
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
![Page 4: Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a.](https://reader036.fdocuments.in/reader036/viewer/2022072011/56649e0c5503460f94af55d4/html5/thumbnails/4.jpg)
The Big Picture
• The inserted gene is transcribed and translated using the RNA Polymerase, ribosomes and other resources in the cell
![Page 5: Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a.](https://reader036.fdocuments.in/reader036/viewer/2022072011/56649e0c5503460f94af55d4/html5/thumbnails/5.jpg)
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
chromosomal DNA
plasmids
![Page 6: Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a.](https://reader036.fdocuments.in/reader036/viewer/2022072011/56649e0c5503460f94af55d4/html5/thumbnails/6.jpg)
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
![Page 7: Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a.](https://reader036.fdocuments.in/reader036/viewer/2022072011/56649e0c5503460f94af55d4/html5/thumbnails/7.jpg)
Transformation
• Process in which foreign DNA is physically inserted into host E. coli cells.
• E. coli that contains recombinant plasmid = Transformed cell Image taken with out permission from
http://summerschool.at/static/irismaria.schoenbrunner/images/transformation.png
![Page 8: Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a.](https://reader036.fdocuments.in/reader036/viewer/2022072011/56649e0c5503460f94af55d4/html5/thumbnails/8.jpg)
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
Image taken without permission fromTransformation Animation. Available at http://www.dnai.org/b/index.html
![Page 9: Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a.](https://reader036.fdocuments.in/reader036/viewer/2022072011/56649e0c5503460f94af55d4/html5/thumbnails/9.jpg)
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
![Page 10: Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a.](https://reader036.fdocuments.in/reader036/viewer/2022072011/56649e0c5503460f94af55d4/html5/thumbnails/10.jpg)
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
![Page 11: Genetic Engineering What is Genetic Engineering? Genetic Engineering = inserting a foreign gene of interest into a host to transcribe and translate a.](https://reader036.fdocuments.in/reader036/viewer/2022072011/56649e0c5503460f94af55d4/html5/thumbnails/11.jpg)
E. Coli as a host
• E. coli is a good host because:– Reproduce quickly (once every 20 minutes)– Nonpathogenic (the strain we use is not
harmful)– Genome fully characterized (all genes have
been sequenced)