Iowa State University of Science and Technolo ?s, Iowa 5001 1 
Department of Genetics 
8 Curtiss Hall 
Telephone: 515-294-3908 
February 10, 1978 
Dr. Donald Fredrickson 
N.I.H. 
9000 Rockville Place 
Bethesda, Maryland 20014 
Dear Dr. Frederickson: 
I have been in communication with Dr. William Garland and others on the 
Advisory Committee on DNA Regulation regarding containment facilities necessary 
for certain proposed recombinant DNA experiments. I have been told by all I 
have contacted that the Guidelines, even the recently revised version, do not 
mention the type of experiment we propose and thus give us no indication of 
the facilities needed. Dr. Garland suggested I write to you pointing out this 
gap in the Guidelines in the hope that some thought might be given to the 
problem before the Committee meets again in April. 
The experiment in question is one involving the isolation and purification 
of messenger RNA's involved in the fixation of nitrogen by Azotobacter 
vinelandii (a non-pathogenic , free living, aerobic bacterium). The use of 
reverse transcriptase is proposed to form the DNA copy, and DNA polymerase to 
restore double stranded structure. This piece of DNA would then be spliced 
into the mitochondrial DNA isolated from either Chlamydomonas reinhardii or 
Solanum tuberosum (potato). Chlamydomonas is a non-pathogenic, free living, 
single-celled, green alga, and the mutant we should be using grows only as 
protoplasts. 
Once spliced into the mitochondrial DNA the DNA would be reintroduced into 
isolated mitochondria which would in turn be reintroduced into their respective 
protoplasts. Growth would be maintained on nitrogen-deficient media and 
nitrogen fixation activity assayed by acetylene reduction activity. 
Our present facilities rate as P2 and could with but small inconvenience 
be transformed into P3. We wish to write up this proposal as soon as possible 
[Appendix A — 234] 
