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Federal Register / Vol. 45, No. 85 / Wednesday, April 30, 1980 / Notices 
7, 1980 meeting, and agreed that the 
proposal should be published in the 
Federal Register for comment. 
The proposal consists of two parts as 
follows: 
A. The following language would be 
substituted for Section IIl-O: 
HI-0 Classification of Experiments Using 
E. coli K-12 and Saccharomyces cerevisiae 
Host-Vector Systems. Most recombinant 
DNA experiments currently being done 
employ E. coli K-12 host-vector systems; 
others employ the S. cerevisiae host-vector 
systems. 
These are the systems for which we have 
the most experience and knowledge. 
Some experiments using E. coli K-12 and S. 
cerevisiae host-vector systems are prohibited 
(see Section I-D). 
Some experiments using E. coli K-12 and S. 
cerevisiae host-vector systems are exempt 
from the Guidelines (see Section I-E). 
Other experiments using E. coli K-12 or 
laboratory strains of S. cerevisiae shall use 
PI physical containment and, except as 
specified in the last paragraph of this section, 
and HV1 host-vector system [i.e. for 
experiments using E. coli K-12 (a) the E. coli 
host shall not contain conjugation-proficient 
plasmids or generalized transducing phages, 
and (b) lambda or lambdoid bacteriophages 
or non-conjugative plasmids [49] shall be 
used as vectors. For experiments in S. 
cerevisiae, laboratory strains shall be used.] 
For these experiments no Memorandum of 
Understanding and Agreement (MUA) as 
described in Section IV-D-l-c need be 
submitted, nor is any registration with NIH 
necessary. However, for these experiments, 
prior to their intiation, investigators must 
submit to their Instutitional Biosafety 
Co mmi ttee (IBC) a registration document that 
contains a description of (a) the source(s) of 
DNA, (b) the nature of the inserted DNA 
sequences, and (c) the hosts and vectors to be 
used. This registration document must be 
dated and signed by the investigator and filed 
only with the local IBC. The IBC shall review 
all such proposals but such review is not 
required prior to intiation of experiments. An 
exception, however, which does require prior 
review and approval by the IBC is any 
experiment in which there is a deliberate 
attempt to have the E. coli K-12 or S. 
cerevisiae efficiently express as a protein 
product the information carried in any gene 
derive from a eukaryotic organism or from 
any virus or viroid which infects a eukaryotic 
organism. 
Experiments involving the insertion into E. 
coli K-12 of DNA from prokaryotes that 
exchange genetic information with E. coli by 
known physiological processes will be 
exempted from these Guidelines if they 
appear on the "list of exchangers” set forth in 
Appendix A (see Section I-E-4). 
For those not on the Appendix A list but 
which exchange genetic information (35) with 
E. coli, experiments may be performed with 
any E. coli K-12 vector (e.g. conjugative 
plasmid). When a nonconjugative vector is 
used, the E. coli K-12 host may contain 
conjugation-proficient plasmids, either 
autonomous or integrated, or generalized 
transducing phages. 
B. The language ”*HVl — Unmodified 
laboratory strains of Saccharomyces 
cerevisiae" would be deleted from 
Appendix D. 
5. Request for Permission to 
Transform Sacchromycopsis lipolytica 
with E. coli/S. cerevisiae Hybrid 
Plasmids. Dr. David M. Ogrydziak of the 
University of California, Davis, requests 
permission to attemt to transform the 
yeast Sacchromycopsis lipolytica with 
defined Escherichia coli / 
Saccharomyces cerevisiae hybrid 
plasmids and to use the hybrid plasmids 
for shotgunning S. lipolytica DNA in E. 
coli and returning the DNA to 5. 
lipolytica. 
6. Proposal to Amend Section II1-B-3 
of the Guidelines. Dr. Allan Campbell of 
Stanford University has proposed that 
the RAC consider amending Section III— 
B-3 of the Guidelines, which currently 
reads as follows: 
m-B-3. Non-HVl systems. Containment 
levels for other classes of experiments 
involving non-HVl systems may be approved 
by the Director, NIH. (See Sections IV-E-1- 
b— (1)— (b). IV-E-l-b-(2)-{c), and IV-E-l-b- 
(3Hb). 
In those cases where genetic exchange has 
not been demonstrated between two 
bacterial species A and B, neither of which is 
known to be pathogenic for man, animals or 
plants, recombinant DNA experiments 
involving only A and B can be conducted 
under P3 containment. (2A) 
Dr. Campbell writes that the P3 
containment level was assigned because 
of apprehensions that the category might 
contain some experiments for which 
that level was appropriate rather than 
that all experiments in this category 
were perceived as requiring P3 
containment. His proposed amendments 
are “intended to assert RAC’s intention 
to review individual proposals in an 
open-minded manner with respect to 
realistic evaluation of possible hazard, 
and to discourage the inference that the 
P3 level, as it applies to a specific 
experiment, has been set on the basis of 
information about the organisms 
involved.” 
Dr Campbell proposes changes in the 
Guidelines as follows: 
A. Section m-B-3, paragraph 2, would 
be amended to read: 
“In those cases when genetic exchange has 
not been demonstrated between two 
bacterial species A and B, neither of which is 
known to be pathogenic for man, animals or 
plants, recombinant DNA experiments 
involving only A and B can be conducted 
under P3 containment [2A]. Lower levels of 
physical containment may be assigned by 
NIH for specific donor-recipient 
combinations. (See Section IV-E-l-b-2-f)." 
B. A new section IV-E-l-b-2-(f) 
would be added to the Guidelines as 
follows: 
”IV-E-l-b-2-{f). Assigning containment 
levels for experiments in which both donor 
and recipient are non-pathogenic 
prokaryotes. (See Section III-B-3)." 
An alternate amendment of the 
second paragraph of Section m-B-3 of 
the Guidelines has been proposed by Dr. 
Irving Johnson of Lilly Research 
Laboratories, as follows (amended 
language in italics): 
"In those cases where genetic exchange 
has not been demonstrated between two 
bacterial species A and B neither of which is 
known to be pathogenic for man, animals or 
plants, recombinant DNA experiments 
involving only A and B can be conducted 
under physical containment and conditions 
specified by the local IBC based on its 
consideration of the biological properties of 
the organisms. " 
7. Request to Delete Language 
Requiring the use of Cauliflower Mosaic 
Virus (CaMV) mutants lacking the 
Aphid Transmission Factor from 
Section III-C-3. Request for Permission 
to Transfer DNA fragments from Aphid 
Transmissible to Aphid 
Nontransmissible CaMV. Dr. Robert J. 
Shepherd of the University of California 
at Davis has requested that the 
following paragraph be deleted from 
section III-C-3 of the Guidelines: 
“The CaMV strain used as a cloning vector 
shall be a mutant that lacks the aphid 
transmission factor.” 
In addition, Dr. Shepherd requests 
that he be permitted to transfer DNA 
segments from aphid transmissible to 
non-aphid transmissible strains of 
cauliflower mosaic virus in order to 
study the factors determining aphid 
transmissibility. This experiment could 
be considered as being prohibited under 
Section I-D-3 of the Guidelines. Dr. 
Shepherd suggests that it is inconsistent 
to restrict studies with cauliflower 
mosaic virus to non-insect transmissible 
strains when no such restriction is 
imposed on studies with bean golden 
mosaic and related viruses, the only 
other known group of DNA plant 
viruses. Dr. Shepherd suggests that 
under the PI containment conditions 
specified in the Guidelines, cauliflower 
mosaic virus is easily confined to a 
growth chamber or greenhouse. In 
addition, the virus has a very restricted 
host range and is neither seed nor pollen 
transmitted, providing even further 
containment. 
8. Proposal to Amend Section 1II-C-4 
of the Guidelines. Dr. Milton Zaitlin of 
Cornell University proposed that 
Section III— C— 4 of the Guidelines be 
amended to redefine a PI greenhouse or 
growth chamber when certain infectious 
agents other than viruses are used as 
vectors. He proposes that the following 
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