Question

You are studying the genetic basis of a blue/orange throat color polymorphism in river guppies. You discover a gene that has an apparent nonsense mutation in orange fish compared to blue ones and this gene associates perfectly with phenotype. Describe an experiment that would functionally determine whether or not this gene is truly involved in production of throat color patches in guppies.

Answer 1

Answer:

Certainly, since you predict a specific gene to be involved in producing throat color patches in guppies, a gene target approach is perfect in determining its function. In this experiment you either knockdown or over express the gene through vector transfection procedures followed by observation in change in throat color.

Steps involved are as follows: (This is only for example, it might alter based on various things)

Isolate specific gene of interest. Cut with restriction enzyme of gene and vector and insert the gene. Make sure the insert contains bacericidal resisitant marker at site.

Incorporate these vectors through electroporation into cells carrying an antibiotic senstive marker.

Next, transfected cells can be selected based on the antibiotic sensitive marker.

Only cells transfected can survive and sensitive cells with no resistant marker, do not survive.