Federal Register / Vol. 46, No. 233 / Friday, December 4 , 1981 / Notices 
59369 
Appendix C — Exemptions Under I-E-5 
Appendix D — (Deleted] 
Appendix E — (Deleted) 
Appendix F — Certified Host-Vector Systems 
Appendix G — Containment Conditions for 
Cloning of Genes Coding for the 
Biosynthesis of Toxins for Vertebrates 
Appendix H — (Deleted] 
I. Scope of the Guidelines 
1-A. Purpose. The purpose of these 
Guidelines is to specify practices for 
constructing and handling (i) 
recombinant DNA molecules and (ii) 
organisms and viruses containing 
recombinant DNA molecules. 
Adherence to those standards by all 
laboratories using recombinant DNA is 
recommended. 
' 1-B. Definition of Recombinant DNA 
Molecules. In the context of these 
Guidelines, recombinant DNA molecules 
are defined as either (i) molecules which 
are constructed outside living cells by 
joining natural or synthetic DNA 
segments to DNA molecules that can 
replicate in a living cell, or (ii) DNA 
molecules that result from the 
replication of those described in (i) 
above. 
1-C. [Deleted] 
I-D. [Deleted] 
I-E. Exemptions. The following 
recombinant DNA molecules are exempt 
from these Guidelines: 
1-E-l. Those that are not in organisms 
or viruses. [5] 
l-E-2. Those that consist entirely of 
DNA segments from a single 
nonchromosomal or viral DNA source, 
though one or more of the segments may 
be a synthetic equivalent. 
l-E-3. Those that consist entirely of 
DNA from a prokaryotic host, including 
its indigenous plasmids or viruses, when 
propagated only in that host (or a 
closely related strain of the same 
species] or when transferred to another 
host by well established physiological 
means; also those that consist entirely of 
DNA from a eukaryotic host, including 
its chloroplasts, mitochondria, or 
plasmids (but excluding viruses), when 
propagated only in that host (or a 
closely related strain of the same 
species). 
l-E-4. Certain specified recombinant 
DNA molecules that consist entirely of 
DNA segments from different species 
that exchange DNA by known 
physiological processes, though one or 
more of the segments may be a synthetic 
equivalent. A list of such exchangers 
will be prepared and periodically 
revised by the Director, NIH, with 
advice of the RAC, after appropriate 
notice and opportunity for public 
comment. (See Section IV-E-l-b-(l)- 
(d).) Certain classes are exempt as of 
publication of these Revised Guidelines. 
The list is in Appendix A. An updated 
list may be obtained from the Office of 
Recombinant DNA Activities, National 
Institutes of Health, Bethesda, Maryland 
20205. 
l-E-5. Other classes of recombinant 
DNA molecules, if the Director, NIH, 
with advice of the RAC, after 
appropriate notice and opportunity for 
public comment, finds that they do not 
present a significant risk to health or the 
environment. (See Section IV-E-l-b- 
(l)-(d).) Certain classes are exempt as of 
publication of these Revised Guidelines. 
The list is in Appendix C. An updated 
list may be obtained from the Office of 
Recombinant DNA Activities: National 
Institutes of Health, Bethesda, Maryland 
20205. 
I-F. General Definitions. See Section 
IV-C. 
II. Containment 
Effective biological safety programs 
have been operative in a variety of 
laboratories for many years. 
Considerable information, therefore, 
already exists for the design of physical 
containment facilities and the selection 
of laboratory procedures applicable to 
organisms carrying recombinant DNAs. 
(6-19) The existing programs rely upon 
mechanisms that, for convenience, can 
be divided into two categories: (i) A set 
of standard practices that are generally 
used in microbiological laboratories, 
and (ii) special procedures, equipment, 
and laboratory installations that provide 
physical barriers which are applied in 
varing degrees according to the 
estimated biohazard. 
Experiments on recombinant DNAs, 
by their very nature, lend themselves to 
a third containment mechanism — 
namely, the application of highly 
specific biological barriers. In fact, 
natural barriers do exist which limit 
either (i) the infectivity of a vector, or 
vehicle, (plasmid or viiiis) for specific 
hosts or (ii) its dissemination and 
survival in the environment. The vestors 
that provide the means for replication of 
the recombinant DNAs and/or the host 
cells in which they replicate can be 
genetically designed to decrease by 
many orders of magnitude the 
probability of dissemination of 
recombinant DNAs outside the 
laboratory. 
As these means of containment are 
complementary, different levels of 
containment appropriate for 
experiments with different recombinants 
can be established by apply various 
combinations of the physical and 
biological barriers along with a constant 
use of the standard practices. We 
consider these categories of 
containment separately here in order 
that such combinations can be 
conveniently expressed in the 
Guidelines. 
In constructing these Guidelines, it 
was necessary to define boundary 
conditions for the different levels of 
physical and biological containment and 
for the classes of experiments to which 
they apply. We recognize that these 
definitions do not take into account all 
existing and anticipated information on 
special procedures that will allow 
particular experiments to be carried out 
under different conditions than 
indicated here without affecting risk. 
Indeed, vye urge that individual 
investigators devise simple and more 
effective containment procedures and 
that investigators recommend changes 
in the Guidelines to permit their use. 
II-A Standard Practices and Training. 
The first principle of containment is a 
strict adherence to good microbiological 
practices. (6-15) Consequently, all 
personnel directly or indirectly involved 
in experiments on recombinant DNAs 
should receive adequate instruction. 
This should, as a minimum, include 
instructions in aseptic techniques and in 
the biology of the organisms used in the 
experiments, so that the potential 
biohazards can be understood and 
appreciated. 
Any research group working with 
agents with a known or potential 
biohazard should have an emergency 
plan which describes the procedures to 
be followed if an accident contaminates 
personnel or the environment. The 
principal investigator should ensure that 
everyone in the laboratory is familiar 
with both the potential hazards of the 
work and the emergency plan. If a 
research group is working with a known 
pathogen where there is an effective 
vaccine it should be made available to 
all workers. Where serological 
monitoring is clearly appropriate it 
should be provided. 
II-B Physical Containment Levels. 
The objective of physical containment is 
to confine organisms containing 
recombinant DNA molecules, and thus 
to reduce the potential for exposure of 
the laboratory worker, persons outside 
of the laboratory, and the environment 
to organisms containing recombinant 
DNA molecules. Physical containment is 
achieved through the use of laboratory 
practices, containment equipment, and 
special laboratory design. Emphasis is 
placed on primary means of physical 
containment which are provided by 
laboratory practices and contaimnent 
equipment. Special laboratory design 
provides a secondary means of 
protection aganist the accidental release 
of organisms outside the laboratory or to 
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