Federal Register / Vol. 45, No. 147 / Tuesday, July 29, 1960 / Notices 
50529 
be free of harmful sequences (see Section III- 
A-3-bJ. It should be clear, however, that the 
funding agency must be notified of IBC 
approvals even when prior agency approval 
is not required. See the Administrative 
Practices Supplement for further discussion. 
No comments were received during 
the thirty-day comment period. These 
proposed changes of the Guidelines 
were considered by the RAC at its June 
5-6, 1980 meeting. The RAC 
recommended the changes by a vote of 
fifteen in favor, none opposed, and no 
abstentions. 
I accept the proposed changes. 
I-N. Request for Permission To 
Transform SaccharomycQpsis Lipolytica 
With E. Coli/S. Cerevisiae Hybrid 
Plasmids 
In response to a request to approve 
the transformation of the yeast 
Saccharomycopsis lipolytica with 
defined E. coli/Saccharomyces 
cerevisiae hybrid plasmids the following 
notice appeared in the April 30, 1980 
Federal Register: 
“Dr. David M. Ogrydziak of the University 
of California, Davis, requests permission to 
attempt 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 die DNA to S. 
lipolytica. 
During the thirty-day comment period 
there were no comments. The RAC 
discussed the proposal during its June 5- 
6, 1980 meeting. It was noted that S. 
lipolytica is not a pathogen, and does 
not colonize animals. A Motion to 
approve these experiments at the Pi 
level of containment was passed by a 
vote of fifteen in favor, none opposed, 
and one abstention. 
I accept this recommendation. 
1-0 Request for Use of Conjugative 
Plasmids and Transducing Phages in an 
Escherichia Coli K-12 Host. 
The following notice was published in 
the Federal Register of April 30, 1980: 
Dr. A. I. Bukhari of the Cold Spring Harbor 
Laboratory has requested permission from 
the RAC to employ conjugative plasmids or 
transducing phages in recombinant DNA 
experiments involving Escherichia coli DNA 
when a small fragment of heterologous DNA 
is used as a linker. In the specific request, Dr. 
Bukhari requests approval to use a fragment 
of Adenovirus 2 DNA as linker DNA. 
No comments were received during 
the thirty-day comment period. 
In the RAC discussion of the request 
at its June 5-6, 1980 meeting, it was 
noted that the small Adenovirus DNA 
linker segments of 1 kb size were not 
derived from the transforming region 
and could, therefore, be treated as inert 
DNA. The motion recommending 
approval of this request passed by a 
vote of fourteen in favor, none opposed, 
and one abstention. 
I accept this recommendation. 
II. Summary of Major Actions Under 
Guidelines 
II-A. Amendment of Section I-D-l 
Section I-D-l is amended to read: 
I-D-l. Formation of recombinant DNAs 
derived from pathogenic organisms classified 
[1] as Class 4 or 5 or from cells known (2A) to 
be infected with such agents, regardless of 
the host-vector system used. 
II-B. Deletion of Section I-D-3 
Section I-D-3 is deleted. Substitute for 
the current wording, the word "deleted." 
II-C. Amendment of Section I-D-6 of the 
Guidelines 
Delete the second paragraph of 
Section I-D-6, which reads as follows: 
We differentiate between small- and large- 
scale experiments with organisms containing 
recombinant DNAs because the probability 
of escape from containment barriers normally 
increases with increasing scale. 
II-D. Amendment of Section I-E 
Section I-E is amended to read: 
I-E. Exemptions. It must be emphasized 
that the following exemptions [4] are not 
meant to apply to experiments described in 
Sections I-D-l to I-D-5 as being prohibited. 
In addition, any recombinant DNA molecules 
involving DNA from Class 3 organisms [1] or 
cells known to be infected with these agents, 
or any recombinant DNA molecules which 
increase the virulence and host range of a 
plant pathogen beyond that which occurs by 
natural genetic exchange, are not exempt 
unless specifically so designated by N1H 
under Section I-E-5. 
II-E. Amendment of Section III 
After the last sentence of Section HI, 
"The use of higher levels of biological 
containment * * * for the purposes of 
the experiment," the following new 
paragraph is to be added: 
Experiments involving recombinant DNA 
from Class 3 organisms [1] or from cells 
known to be infected with these agents may 
be conducted at P3 containment in E. coli K- 
12 EKl hosts (see Section m-O). Containment 
levels for all other experiments with Class 3 
organisms or with recombinant DNA which 
increases the virulence and host range of a 
plant pathogen beyond that which occurs by 
natural genetic exchange will be determined 
by NIH. (See Section IV-E-l-b-2-e). 
II-F. Amendment of Section III-O of the 
Guidelines 
Section III-O of the Guidelines is 
amended to read as follows: [This 
amended language combines (1J the 
language of the proposal to include 
Saccharomyces cerevisiae host-vector 
systems under Section III-O, (2) the 
language of the proposal to include Ff 
phages under Section III-O, and (3) a 
paragraph pertaining to CDC Class 3 
agents and certain plant pathogens.] 
III-O. 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). 
Experiments using E. coli K-12 host-vector 
systems and DNA from Class 3 organisms [1] 
or from cells known to be infected with these 
agents will be conducted at P3 containment 
or at a lower level as specified by NIH (See 
Section IV-3-l-b-2-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, > 
an HV1 hostvector system [i.e., for 
experiments using E. coli K-12 (a) the E. coli 
host shall not contain conjugation-proficient 
plasmids or generalized transducinjg phages, 
and (b) lambda or lambdoid of Ff 
bacteriophages or non-conjugative plasmids 
[49] shall be used as vestors. 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 initiation, 
investigators must submit to their 
Institutional Biosafety Committee (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 initiation 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 efficiently 
express as a protein product the information 
carried in any gene derived from a eukaryotic 
organism or from any virus or viroid which 
infects a eukaryotic organism. 
Experiments involving the insertion into E. fj 
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 Appendix A list but which 
exchange genetic information [35] with E. 
