Test 3 Transgenic and Knockout Animals

Question 1

Why mammary glands?

Answer 1

Milk is a renewable, secreted fluid that is produced in substantial quantities and can be collected frequently without harm to the animal

Question 2

promoters for milk production

Answer 2

• milk protein promoter
• Whey acidic protein promoter
• b-Lactoglobin promoter
• b-Casein promoter

Question 3

The basic process for the creation of a transgenic animal

Answer 3

• isolate embryonic stem cell from animal
• put into tissue culture and grown
• clone DNA recombine into chromosome
• introduce selectible marker -> only cells that take up transgene will survive
• inject transgene into blastosis
• blastosis injected into animal -> animal gives birth and some offsprings will have transgene
• breed until you have homozygous female

Question 4

ATryn

Answer 4

• recombinant antithrombin protein purified from the milk of genetically modified goats
• molecule that inactivates several enzymes of the coagulation system

Question 5

Ruconest

Answer 5

• secreted in the milk of transgenic rabbits

- for Hereditary Angioedema

Question 6

reporter gene

Answer 6

surrogate for the expression of the protein that you’re interested in; you don’t have to kill the animal

Question 7

Green Fluorescent Protein (GFP)

Answer 7

• combine GFP with promoter of interest or
• fuse GFP with coding region of some gene
• shine a light of specific wavelength on animal

Question 8

Why knock out a gene?

Answer 8

• to see what it does

- determine the developmental and physiological consequences of inactivating a particular gene

Question 9

Null allele

Answer 9

a nonfunctional gene generated by either deletion of the entire gene or mutation of the gene resulting in the synthesis of an inactive (non-functional) protein

Question 10

How to knockout a gene

Answer 10

• make recomb DNA molecule
• introduce into cell and select for neomycin resistance
• after homologous recombination, gene knocked out in cell chromosome by replacement with neoR

Question 11

creation of knockout mouse

Answer 11

• isolate embryonic stem cells
• introduce plasmid
• introduce DNA into tissue culture -> cells pick up DNA with neomycin resistance marker
• add antibiotic and only those cells survive
• introduce in blastosis -> insert in mother -> breed until you have homozygous