Attachment II - Page 4 
4 
(2) What is the sequence of the inserted DNA? For example, 
-hat promoters and enhancers are used? Hew was the 
inserted DMA prepared? 
(3) In what media are the vector and inserted DC3V grown or 
maintained? 
(4) How will the stability of the vector and inserted DNA in 
culture be assayed? 
(5) How will the absence of contaminants (e.g., bacteria, 
fungi, viruses) in the culture(s) be demonstrated? 
b. Prior laboratory research 
(1) What laboratory studies have been performed in tissue 
culture? With cells frem which species? What have been 
tiie results of these studies with respect to the following 
points? 
(a) Integration sites of vector and inserted DNA, if any 
(b) Structural stability of vector and inserted DNA, absence 
of other adventitious reccmbinational events, or other 
variant DNAs 
(c) Expression of inserted gene: its expression, stability, 
and regulation (including time and amount of expression) 
[ 112 ] 
