Tutorial 3

This tutorial is intended to strengthen your understanding of gene cloning, PCR and Southern and northern (and even western) blotting. Before the tutorial put some thought into and attempt to answer the questions below. You will need to do some reading either from the web or possibly even from a book! Print out this page or use the sheet handed out in the lectures and bring your answers to the tutorial. Be prepared to discuss them.

terms and expressions used in human molecular genetics:

Come to the tutorial prepared to explain the meanings of the following terms:


1. plasmid clone
2. cDNA library (what considerations are important in the design and construction of such a library?)
3. oligonucleotide
4. genomic library (again, what considerations are important in the design and construction of such a library?)
5. micro array
6. Blot - as in Southern, Northern, Western

(Not sure what any of these terms are? For a first clue look them up in the b241 glossary)

gene cloning

In days gone by, you might have made your reputation as a hot human geneticist by identifying the gene in which mutations cause a particular genetic disease 'a disease gene' e.g. the gene "for" cystic fibrosis.

Now you are more likely to investigate such questions as "What gene products interact with the gene product of the disease gene?".

In the tutorial, be prepared to explain:
1. How would you have identified the gene which codes for a particular protein - let's say carbonic anhydrase III?
2. How would you look for factors regulating carbonic anhydrase III expression?
3. How would you find other proteins with which carbonic anhydrase III interacts?

references

A very clear description of the principles of Southern, northern and western blotting and gene cloning is to be found in the MIT Biology Hypertextbook at this page which may tell you all you need to know to answer the first questions.