1 / 15

Bacterial Transformation

Bacterial Transformation. What is transformation?. Changing the genes and phenotype of a bacteria by uptake of foreign/new DNA Let’s review bacterial DNA first…. Bacterial genome. Bacteria are prokaryotes—no nucleus. The area where DNA is located is called the nucleoid

inicholson
Télécharger la présentation

Bacterial Transformation

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Bacterial Transformation

  2. What is transformation? • Changing the genes and phenotype of a bacteria by uptake of foreign/new DNA • Let’s review bacterial DNA first…

  3. Bacterial genome • Bacteria are prokaryotes—no nucleus. • The area where DNA is located is called the nucleoid • DNA is organized in one double stranded circular molecule

  4. What is carried on the Plasmid? • The plasmid contains genes necessary for survival and can be passed from one bacteria to another • Color Marker gene- Beta-galactosidase-produces enzyme that breaks down lactose • Antibiotic Resistance: Some bacteria have genes coding for enzymes that destroy certain antibiotics!

  5. pBLU plasmid B-galactosidase gene Ampicillin resistance gene

  6. The transformation lab… • Our plasmid—pBlu plasmid • Into E. coli (scary?…no!) • Our plasmid contains genes for: • amp= ampicillin (an antibiotic) resistance • Beta-galactosidase-an enzyme that converts X-Gal  Indo Blu RNA Enzyme that breaks down X-Gal to make Indo- Blu RNA Protein that allows for antibiotic resistance

  7. How do we get the plasmid inside of the bacteria? 2. pBlu plasmid • Obtain E. Coli bacteria cells + Add to CaCl2 (helps plasmid attach to bacteria) • Add plasmid to same microtube 1. E. Coli

  8. How do we get the plasmid inside the bacteria? • Wait…and then 3. Heat shock! This temporarily opens pores to allow the plasmid to enter the bacteria…timing is critical!!!

  9. Growing the bacteria • After they have received the plasmid… • Placed on a growth media and allowed to grow.

  10. How will we know if the bacteria actually got the plasmid?? • Any ideas? • We can grow the bacteria on a plate: • That contains ampicillin and X-Gal • Regular bacterial medium • What do you predict will happen in each?

  11. pBlu (+/-plasmid) pBlu (+ plasmid) pBlu (- plasmid) pBlu (- plasmid) pBlu (- plasmid) Predict What will we observe??? LB LB/AMP LB/AMP LB/AMP/X-gal LB/AMP/X-gal

More Related