Attachment IV - Page 26 
-4- DRAFT 
NOVEMBER 7, 1983 
III. Expression Systems 
Recombinant DMA technology Involves systematically arranging and manipulating 
specific segments of nucleic acid to produce a composite molecule which, 
when placed Into an appropriate host environment, will yield a desired product. 
There are three general methods for obtaining a specific coding segment: (a) 
reverse transcription of mRNA to complementary DNA; (b) Isolation of genomic 
DNA or RNA; or (c) chemical synthesis as predicted from a peptide sequence. 
The manufacturer should provide a description of the method used to prepare 
the segment coding for the desired product, Including both the cell type and 
origin of the source material, a detailed nucleotide sequence analysis and a 
restriction enzyme digestion map of the cloned segment. If a cloned polynucleotide 
contains more Information than the coding sequence, l.e. Introns or flanking 
sequences, then these additional sequences should be Identified. 
The construction of the vector used for expression of the cloned nucleotide 
segment into its respective product should also be thoroughly described. This 
description should Include a detailed explanation of the source and function of 
the component parts of the vector e.g. origins of replication, antibiotic 
resistance genes, promoters, enhancers, whether or not the product is being 
synthesized as a fusion protein. The restriction enzyme digestion map of the 
entire constructed vector should be provided. 
The host cell system which will generate the product Is coordinated to 
fit the expression vector. It is, therefore, important that a complete 
description of the source, relevant phenotype and genotype of the host be provided. 
If the host cell is of mammalian origin then data concerning its 
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