Dr. Donald S. Fredrickson 
September 23, 1978 
Page 6 
d. Another problem not considered concerns the fact that the exclusion 
granted in Section I-E-4 allows one to clone, for example. Salmonella 
DNA into Pseudomonas or vice versa even though there are no data to 
indicate that Salmonella species can exchange chromosomal genetic in- 
formation with Pseudomonas species. In other words, the list of 
organisms in Appendix A (p. 33089) is based almost entirely on the 
ability of organisms on the list to transfer chromosomal information 
to £. col i K-12. Thus, if it is the intent to conclude that if both 
organisms A and B can exchange chromosomal information with organism 
C that exchange between organisms A and B would occur even though this 
had not been demonstrated, this should be explicitly stated. I might 
add that such an assumption becomes somewhat farfetched when one con- 
siders that Acinetobacter calcoaceticus has a DNA guanine-cytosine con- 
tent of 42-43% and another organism listed in Appendix A, Rhodopseudo - 
monas sphaeroides , has a DNA guanine-cytosine content of 71%. 
e. Another problem with Appendix A is that it is incomplete. Lists of ex- 
changers in the genera Bacillus , Streptococcus , Haemophilus and Strepto - 
myces are lacking and these omissions will impede recombinant DNA re- 
search with these organisms and also the development of new host-vector 
systems. This is also bothersome since although it is now possible, for 
example, to clone Bacillus pumilis DNA in EL subtil is , the Proposed 
Revised Guidelines prohibit such an experiment until such time as the RAC 
approves certain EL subtil is derivatives as HV1 or resolves the problem 
in some other way. 
7. Section II. Containment (p. 33070-33076) . Overall, this section is extremely 
well done. I very much support the idea of alternate practices of achieving physical 
containment at the P3 and P4 level and believe that the Laboratory Safety Monograph 
is extremely well done and will be most helpful and an important improvement over 
Appendix D of the Current NIH Guidelines. I do, however, have some minor points as 
enumerated below. 
8. Section 1 1 -B-2-a- ( 13 ) . dealing with laboratory gowns, coats or uniforms (p. 
33072). Although it should be obvious that laboratory clothing is designed to pro- 
tect undergarments from contamination with biological materials and that investiga- 
tors wearing such protective clothing are most likely to experience aerosolized or 
spilled materials on their fronts, I have noted in laboratories that I have visited 
as well as in my own laboratory that investigators have a tendency to leave such 
laboratory coats open. I thus think it might be helpful to be more explicit in 
this section and to state that such laboratory gowns, coats or uniforms, etc. should 
be buttoned, zippered, etc. 
9. Section II-B-2-b. Containment Equipment (p. 33072). The use and necessity of 
biological safety cabinets in P2 laboratories is ambiguous. This section refers to 
aerosol-producing equipment but makes no mention of aerosol -producing procedures. 
It is also noteworthy that most of the equipment listed except for centrifuges is 
seldom used in recombinant DNA research. The most prevalent manipulations in labo- 
ratories using recombinant DNA technology with microorganisms are the growth of cul- 
[A-313] 
