back to Molecular Biology Homepage

back to Objectives and Outlines Page

Eukaryotic Transcriptional Regulation, Part I

Oct 25, 2000

---

Announcements:

• Week 6 Lab Reports are graded, and a key is online
• Homework #5 is graded, and its key is online
• I made new pCITE vector stocks (both 2a and 4a) yesterday--both will be available at a concentration of 0.250 mg/ml for use as controls in today's lab. I'd suggest using 250 ng per digest.
• Homework #7 (on Jacob et al) will go online tonight--keep an eye out for it

Learning objectives for Wed 10/25

Students should be able to…

• 1. Compare and contrast prokaryotic and eukaryotic transcriptional regulation. Include a discussion of the following factors: distance of "cis" elements from the transcription start site, number of control elements involved in a particular gene, assembly of an initiation complex, and modulation of chromatin structure.

  2. Describe how the question of regulation of a particular eukaryotic gene could be addressed. Explain the types of experiments that could be used and a logical order in which to pursue them. For each type of experiment, describe the kind of information that could be obtained and a reason why such information could be useful.

  3. (If we have time to get to this) Explain how the yeast two-hybrid system can be used to identify protein-protein interactions. Describe how this system was developed from the study of a yeast transcriptional activator, Gal4.

   

Outline:

• Studying transcription in eukaryotes: why it matters
• Mapping transcriptional start sites--a way to begin your analysis
• Promoter-reporter fusions and "promoter bashing" experiments--identifying control regions
• Isolating and identifying proteins that bind to control regions
• An aside…the yeast two-hybrid system

back to Objectives and Outlines Page

back to Molecular Biology Homepage