1 / 24

Modeling Plasmid Selection

Modeling Plasmid Selection. And Authentic Assessment. 6-2010. http://biology2020.wikispaces.com/. With the pGlo Plasmid. Bacteria are very useful organisms in genetic engineering. They are able to bring in plasmids. Into which DNA of interest may be added. The Problem?.

trang
Download Presentation

Modeling Plasmid Selection

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. Modeling Plasmid Selection And Authentic Assessment 6-2010

  2. http://biology2020.wikispaces.com/

  3. With the pGlo Plasmid • Bacteria are very useful organisms in genetic engineering. • They are able to bring in plasmids. • Into which DNA of interest may be added.

  4. The Problem? • Finding the bacteria that have taken in the modified plasmids. • You can see the bacteria, but not the plasmid. • Must look for some visible effect of the plasmid.

  5. What to look for? • Growing in the presence of antibiotic. • The effect of a gene, like the glow of pGLO

  6. Cut Four Paper Ovals(or…draw 4 ovals on a large sheet of paper.) • Each represents a bacterial cell

  7. Make Four Chromosomes • Out of four chenille stems • Bacterial • Circular • Give a little twist to prevent confusion with a nucleus. • Add to “cells.” • (Or just draw this chromosome in on paper)

  8. Form your plasmidsUse 1/2 a chenille stem each. Make two plasmids as below. Twist into ring but leave tails Regulatory Gene Ara B Ara A AraD bla Ara C Structural Genes Antibiotic Resistance GFP Bla codes for beta lactamase, a protein which confers antibiotic resistance. By the way…great place to teach the operon. Reserve

  9. Restriction Enzymes • Plasmids are cut with the same restriction enzyme used to cut the DNA to be inserted. A restriction enzyme which leaves overhanging sticky ends is needed for this this procedure. This provides the free base pairs needed to combine the plasmid DNA with the source DNA. Restriction Enzyme Cut from EcoRI

  10. Procedure: • Open the plasmids you prepared by untwisting the ½ chenille stem. • This represents the recognition site for the restriction enzyme. Remove ara B, araA, and araD (last 4 beads). • Add a green bead to represent the GFP (gene of interest). • GFP shares the same sticky ends as the plasmid.

  11. Modeling pGlo LB/AMP LB LB/AMP LB/AMP/ARA -pGLO -pGLO +pGLO +pGLO

  12. Bacteria Lives? LB/AMP LB LB/AMP LB/AMP/ARA Yes No Yes Yes -pGLO -pGLO +pGLO +pGLO

  13. Bacteria Glows? LB/AMP LB LB/AMP LB/AMP/ARA No No No Yes -pGLO -pGLO +pGLO +pGLO

  14. Great Lab for Authentic Assessment Let’s try it.

  15. More Authentic Assessment • Is this gel in the chamber correctly? • How do you know?

  16. 2nd block may get this!

  17. Provide a ruler and some semi-log paper. Ask: How big is fragment X? Which is larger , X or Y?

  18. Micropipettor • Have out a micropipettor and a microtube. • Instructions: • Pipet 50 µL of water into the microtube. • Close and write your initials on the top with the permanent marker. • Place in the box by clicker number. • Set the micropipettor to the day of the month you were born on.

  19. Osmosis • Provide a scale and weigh boat. Just use water…but ask… • This bag has been in this beaker of distilled water for 30 minutes. • The initial mass of the bag was 30 g. • Is the bag hypertonic, hypotonic, or isotonic to the beaker? • Which way did water move?

  20. Fruit Flies Give sex and eye type.

  21. Mitosis Identify the stage of mitosis at the tip of the pointer.

  22. Respiration Read the pipette.

  23. References • Biotechnology Explorer, pGLO Bacterial Transformation Kit. BioRad.

  24. http://www3.bio-rad.com/images/pglomap2.gif

More Related