Federal Register / Vol. 45, No. 147 / Tuesday, July 29, 1980 J Notices 
50525 
however, is reference to RUB 331 which 
has been certified as the host 
component of B. subtilis HVl systems. 
Added to Appendix E is a description of ' 
certified EK2 and HV2 E. coli and S. 
cerevisiae systems. 
I-B. Proposal To Amend Section III-O of 
the Guidelines To Permit Use of Ff 
Phages 
In January 1980 (45 FR 8724) the 
National Institutes of Health (NIH) 
promulgated revised Guidelines. The 
revised Guidelines included a Section, 
III-O, "Classification of Experiments 
Using the E. coli K-12 Host-Vector 
Systems,” which reads, in part, as 
follows: 
* * * experiments using E. coli K-12 shall 
use PI physical containment and * * * an 
EKl host-vector system [i.e., (a) the host shall 
not contain conjugation-proficient plasmids 
or generalized transducing phages and (b) 
lambda or lambdoid bacteriophages or 
nonconjugative plasmids shall be used as 
vectors].* * * 
No reference to the use of Ff 
bacteriophages (filamentous single- 
strand male-specific bacteriophages 
such as M13 and fd) was made in 
Section III-O. Consequently, when the 
revised Guidelines were promulgated in 
January 1980, investigators were 
informed that Ff bacteriophages might 
continue to be used as a component of 
EKl systems at the containment levels 
specified by the 1978 Guidelines. 
In December 1979, the RAC was asked 
to consider how E. coli K-12 host-vector 
systems employing Ff phages should be 
treated under the 1980 Guidelines. In 
response to this NIH request, the Host- 
Phage Subcommittee drafted a 
recommendation which was presented 
to the full RAC at the March 6-7, 1980 
meeting. The RAC agreed that the 
proposed amended language of Section 
III-O should be published in the Federal 
Register for thirty days of comment. 
Accordingly, the following language 
appeared in the Federal Register of April 
30, 1980 [45 FR 28904]: 
* * * experiments using E. coli K-12 shall 
use Pi physical containment and, * * * an 
EKl host-vector system [i.e., (a) the host shall 
not contain conjugation-proficient plasmids 
or generalized transducing phages and (b) 
lambda or lambdoid or Ff bacteriophages or 
non-conjugative plasmids shall be used as 
vectors].* * * 
No comments on this proposal were 
received during the thirty-day comment 
period. 
At the June 5-6, 1980 meeting, the 
RAC once again considered this issue. 
The safety of Ff phages was discussed. 
By a vote of fourteen in favor, none 
opposed, the RAC recommended that 
the Host-Phage Subcommittee proposal 
to include Ff phages under Section III-O 
be accepted. 
I accept this recommendation. 
I-C. Proposed Changes To Allow 
Recombinant DNA Experiments With 
Class 3 Organisms and Certain Plant 
Pathogens 
In December 1979, the National 
Institute of Allergy and Infectious 
Diseases (NIAID) requested that the 
NIH consider whether, and under what 
conditions, recombinant DNA 
technology might be used to study CDC 
Class 3 etiological agents. Currently 
Section I-D-l of the Guidelines 
prohibits: 
Formation of recombinant DNAs derived 
from the pathogenic organisms classified [1] 
as Class 3, 4 or 5 [2] or the cells known [2A] 
to be infected with such agents, regardless of 
the host-vector system used. 
This issue was presented to the RAC 
at its December 6-7, 1979 meeting and a 
working group was appointed to study 
the issue. A draft proposal which would 
revise the Guidelines so as to remove 
Class 3 agents from Section I-D-l of the 
Guidelines was subsequently presented 
to the full RAC at its March 6-7, 1980 
meeting. 
At the March 6-7, 1980 meeting, a 
request from Dr. Clarence Kado of the 
University of California, Davis, to delete 
Section I-D-3 of the Guidelines was also 
presented to the RAC. Section I-D-3 
prohibits: 
Deliberate creation by the use of 
recombinant DNA of a plant pathogen with 
increased virulence and host range beyond 
that which occurs by natural genetic 
exchange. [2A] 
During the discussion of this issue it 
was>suggested that the mechanism used 
to remove CDC Class 3 agents from 
Section I-D-l of the Guidelines might be 
appropriate for removing recombinant 
DNA experiments involving certain 
plant pathogens from Section I-D-3. 
Accordingly, the proposals have been 
combined. 
The proposed changes would remove 
recombinant DNA experiments with 
CDC Class 3 organisms and certain 
recombinant DNA experiments with 
plant pathogens from the prohibited 
classes of experiments. The proposal 
would permit experiments with CDC 
Class 3 organisms to be conducted at P3 
containment in E. coli K-12 EKl host- 
vector systems. The containment levels 
for other recombinant DNA experiments 
would be assigned by the Director, NIH 
after review by the RAC. 
It is felt that the proposal would 
reduce administrative procedures, 
thereby increasing flexibility and 
encouraging the use of recombinant 
techniques to elucidate the pathogenic 
process. It was reasoned that P3 
containment, which is considered 
adequate for work with the pathogenic 
organisms themselves, should be 
adequate, or more than adequate, for 
work with E. coli K-12 containing 
recombinant DNA from these organisms. 
The proposed amendments to the 
Guidelines appeared in the Federal 
Register of April 30, 1980 [45 FR 28904] 
as follows: 
A. Section I-D-l is to be 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. 
B. -Section I-D-3 is to be deleted. 
C. Section I-E is to be 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 NIH under Section I-E-5. 
D. Section HI. After the last sentence, “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 III-O). Containment ji 
levels for all other experiments with Class 3 j 
organisms or with recombinant DNA which 
increases the virulence and host range of a 
plant pathogen will be determined by NIH 
(See Section IV-E-l-b-2-e). 
E. Section III-O. After the third paragraph, | | 
‘Some experiments * * * are exempt from 
the Guidelines (see Section I-E),’ the 
following new paragraph is to be added: 
Experiments using E. coli K-12 EKl 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). 
F. A new Section IV-E-l-b-2-e is to be 
added as follows: 
IV-E-l-b-2-e. Assigning containment 
levels for experiments with recombinant 
DNA from Class 3 organisms [1] and for 
experiments which increase the host-range 
and virulence of plant pathogens. 
G. Section V, footnote 2, is to be amended 
to read;- 
2. For experiments using Vesicular 
Stomatitis virus (VSV), contact the NIH 
Office of Recombinant DNA Activities. 
H. Section V, footnote 36. The second 
sentence is to be amended to read: 
(As noted in the Prohibition Section, the 
use of viruses classified [1] as Class 4 or 5 is 
prohibited.) 
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