Federal Register / Vol. 47, No. 77 / Wednesday. April 21, 1982 / Notices 
17187 
experiments are specified in Appendix 
F. 
III-A-2. Deliberate release into the 
environment of any organism containing 
recombinant DNA. 
III-A-3. Deliberate transfer of a drug 
resistance trait to microorganisms that 
are not known to acquire it naturally 
[2A], if such acquisition could 
compromise the use of the drug to 
control disease agents in human or 
veterinary medicine or agriculture. 
III-B. Experiments that Require IBC 
Approval Before Initiation. Investigators 
performing experiments in this category 
must submit to their Institutional 
Biosafety Committee fIBC), prior to 
initiation of the experiments, a 
registration document that contains a 
description of: (a) The source(s) of DNA, 
(b) the natme of the inserted DNA 
sequences, (c) the hosts and vectors to 
be used, (d) whether a deliberate 
attempt will be made to obtain 
expression of a foreign gene, and, if so, 
what protein will be produced, and (e) 
the containment conditions specihed in 
these Guidelines. 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 prior to iniliation of the 
experiments. Requests for lowering of 
containment for experiments in this 
category will be considered by NIH. 
(See Section IV-E-l-b-(3).) 
III-B-1. Experiments Using CDC Class 
2, Class 3, Class 4, or Class 5 Agents [1] 
as Host-Vector Systems. 
III-B-l-a. Experiments involving the 
introduction of recombinant DNA into 
CDC Class 2 agents can be carried out 
at P2 containment. 
III-B-l-b. Experiments involving the 
introduction of recombinant DNA into 
CDC Class 3 agents can be carried out 
at P3 contaiiunent. 
UI-B-l-c. Experiments involving the 
introduction of recombinant DNA into 
CDC Class 4 [51] or Class 5 agents can 
be carried out at P4 containment. A 
USDA permit is required for work with 
Class 5 agents [48]. 
III-B-2. Experiments in Which DNA 
from CDC Class 2, Class 3, Class 4, or 
Class 5 Agents [1] is Cloned in 
Nonpathogenic Prokaryotic or tower 
Eukaryotic Host-Vector Systems. 
IIl-B-2-a. Recombinant DNA 
experiments in which DNA from CDC 
Class 2 or Class agents [1] is transferred 
into nonpathogenic prokaryotes or lower 
eukaryotes may be performed under P2 
containment. Specific lowering of 
containment to PI for particular 
experiments can be approved by the 
IBC. Many experiments in this category 
will be exempt from the Guidelines (See 
Sections III-D-4 and III-D-5). 
Experiments involving the formation of 
recombinant DNAs for certain toxin 
genes require RAC review and NIH 
approval (see Section IU-A-1), or must 
be carried out under NIH specified 
conditions as described in Appendix F. 
IlI-B-2-b. Recombinant DNA 
experiments in which DNA fi'om CDC 
Class 4 [51] or Class 5 agents is 
transferred into nonpathogenic 
prokaryotes or lower eukaryotes can be 
performed at P2 contaiiunent after 
demonstration that only a totally and 
irreversibly defective fraction of the 
agent's viral genome is present in a 
given recombinant. In the absence of 
such a demonstration, P4 containment 
should be used. 
Note . — A USDA permit is required for work 
with Class 5 pathogens [48]. 
III-B-3. Experiments Involving the 
Use of Infectious Animal or Plant 
Viruses or Defective Animal or Plant 
Viruses in the Presence of Helper Virus 
in Tissue Culture Systems. 
Note: Recombinant DNA molecules 
which contain less than two-thirds of 
the genome of any eukaryotic virus (all 
virus from a single Family [36] being 
considered identical [50]] may be 
considered defective and can be used, in 
the absence of helper, under the 
conditions specified in Section III-C. 
III-B-3-C, Experiments involving the 
use of infectious CDC Class 2 animal 
viruses [1], or defective CDC Class 2 
animal viruses in the presence of helper 
virus, can be performed at P2 
containment. 
III-B-3-b. Experiments involving the 
use of infectious CDC Class 3 animal 
viruses [1], or defective CDC Class 3 
animal viruses in the presence of helper 
virus, can be carried out at P3 
containment. 
III-B-3-a. Experiments involving the 
use of infectious CDC Class 4 or Class 5 
animal virus [1], or defective CDC Class 
4 or Cass 5 animal viruses in the 
presence of helper virus, may be carried 
out under P4 containment. A USDA 
permit is required for work with CDC 
Class 5 pathogens [48]. 
III-B-3-d. ^periments involving the 
use of infectious animal or plant viruses, 
or defective animal or plant viruses in 
the presence of helper virus, not covered 
by Sections III-B-3-a, III-B-3-b, or III- 
B-3-C may be carried out under PI 
contaiiunent. 
III-B-4. Recombinant DNA 
Experiments Involving Whole Animals 
and Plants. 
III-B-4-a. DNA from any source 
except for greater than one quarter of a 
eukaryotic viral genome may be 
transferred to any non-human 
vertebrate organism and propagated 
under conditions of physical 
containment comparable to Pi and 
appropriate to the organism under study 
[2A]. 
in-B-4-b. For all experiments 
involving whole animals and plants and 
not covered by ni-B-4-a, the 
appropriate containment will be 
determined by the IBC. 
ni-B-5. Experiments Involving More 
Than 10 Liters of Culture. The 
appropriate containment will be decided 
by the IBC. Where appropriate, the 
large-scale containment 
recommendations of the NIH should be 
used (45 FR 24968). 
III-C. Experiments That Require IBC 
Notice Simultaneously With Initiation 
of Experiments. Experiments not 
included in Sections m-A, lU-B, lU-D, 
and subsections of these Sections are to 
be considered in Section ED-C. All such 
experiments can be carried out at PI 
containment. For experiments in this 
category, a registration document as 
described in Section III-B must be dated 
and signed by the investigator andf filed 
with the local IBC. The IBC shall review 
all such proposals, but IBC review prior 
to initiation of the experiment is not 
required. 
For example, experiments in which all 
components derive from non-pathogenic 
prokaryotes and non-pathogenic lower 
eukaryotes fall under Section III-C and 
can be carried out at Pi containment. 
Caution: Experiments Involving 
Formation of Recombinant DNA 
Molecules Containing no more Than 
Two-Thirds of the Genome of any 
Eukaryotic Virus. Recombinant DNA 
molecules containing no more than two* 
thirds of the genome of any eukaryotic 
virus (all viruses from a single Family 
[36] being considered identical [50] may 
be propagated and maintained in cells in 
tissue culture using PI containment. For 
such experiments, it must be shown that 
the cells lack helper virus for the 
specific Families of defective viruses 
being used. If helper virus is present, 
procedures specified under Section III- 
B-3 should be used. The DNA may 
contain fragments of the genome of 
viruses from more than one Family but 
each fragment must be less than two- 
thirds of a genome. 
III-D. Exempt Experiments. The 
following recombinant DNA moleculses 
are exempt from these Guidelines and 
no registration with the IBC is 
necessary. 
lU-D-l. Those that are not in 
organisms or viruses. 
III-D-2, Those that consist entirely of 
DNA segments from a single 
nonchromosomal or viral DNA source. 
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