Question 12: 

Your results of the mouse lung study demonstrate that expression of Vegf (Vascular endothelial growth factor) is restricted to the posterior region of the lung epithelial buds.  Your supervisor is very excited about your findings and it is decided that a lung specific Vegf knockout should be generated using a mouse model.  

Explain the process by which you would generate such a knockout (ie. the steps involved). Use a diagram.  

[10 marks; 2 marks for overall diagram, and 2 marks each for  FOUR major steps. Include at the final step what happens genetically.]

(HINT: see Craig’s lecture on mouse models!)

Question 2:  Features of a mouse embryo. [10 marks].  

Take a photo of the mouse embryo. State the embryo age, and label the following features: Forelimb bud, hindlimb bud, forebrain, midbrain, hindbrain, branchial arches, eye, heart, and somites.                                                                                       [10 marks - 1 mark for the image, 9 marks for the labels]

MOUSE PRACTICAL: PART B: 

Question 7:  LacZ stained embryos [6 marks]. 

Take a photo of a stained embryo, showing areas stained with the LacZ solution. NOTE: You may present an image of another embryo if yours does not show LacZ staining.  What major tissues are stained? Label these structures. (6 marks – for correct labelling of at least 3 stained structures; 2 marks each0.

Question 6:  Experimental scenario [10 marks].

b) Imagine you have identified a novel gene using the approach in (a) above. How might you test if this gene has a conserved role in zebrafish vertebral column development?  Without using a diagram, outline a method and the expected outcome.  (4 marks).                           

Question 5: 

What kind of protein/product does this gene encode? 

[2 marks]

Question 4:  [8 marks].

Take a photo of the mutant and upload the labelled image. Using arrows, Identify THREE specific abnormalities that you observe. Include relevant information in a Figure Legend: gene affected, and state the abnormalities .   (8 marks, consisting  of:  the image = 1 mark; labelling THREE abnormalities on the image (2 marks x3 abnormalities); figure legend, statement on gene(s) affected (1 mark). 

Question 3:  Mouse mutant analysis. [5 marks]. 

Using your observations, and the associated paper, identify the mouse mutant model and fill in the table below. (State at least THREE abnormalities).