Factor Vlll

Factor Vlll is a glycoprotein synthesized in liver cells. Many haemophiliacs, who are deficient in Factor Vlll, are now treated by regular injections of genetically engineered Factor Vlll.

Outline how the isolated gene for human Factor Vlll is obtained and inserted into a host cell

Use restriction enzymes to cut the human gene and mammalian cell chromosome

Isolate the human Factor Vlll and mammalian cell and insert the DNA encoding the human factor Vlll into mammallian cells grown in culture.

Human Factor Vlll gene is coupled to the promoter for the ovine(sheep) milk protein beta-lactoglobulin.

State 1 advantage of using recombinant Factor Vlll instead of blood derived Factor Vlll.

Recombinant Factor Vlll does not require injection and thus no risk of infections such as HIV.

Suggest why the host cell used to produce genetically engineered Factor Vlll must be a mammalian cell and not a bacteria cell.

E coli cannot be used as it lacks the machinery to attach carbohydrate properly.
(Human Factor Vlll is a glycoprotein)



Electrical pulse used to introduce activated mammallian cell int enucleated cell.