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Federal Register / Vol. 4B. No. 2.34 / Monday, Decemler 7, 19ttl / Notices 
neurotoxin). Specific approval has been 
given for the cloning in E. colt K-12 of 
ONAs containing genes coding for the 
biosynthesis of toxins which are lethal 
to vertebrates at 100 nanograms to 100 
micrograms per kilogram body weight. 
Containment levels for these 
experiments are specified in Appendix 
G. 
“IlI-A-2. Deliberate release into the 
environment of any organism containing 
recombinant DNA. 
‘‘III-A-3. Deliberate transfer Ok' a drug 
resistance trait to microorganisms that 
are not known to acquire it naturally, if 
such acquistion 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 (IBC), prior to 
initiation of the experiments, a 
registration document that contains a 
description of; (a) The source(s) of DNA, 
(b) the nature 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 specified in 
these Guidelines. This registration 
document must be dated and signed by 
the investigator and Bled only with the 
local IBC. Tlie IBC shall review all such 
proposals prior to initiation 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 * as Host- Vector Systems. 
“IIl-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 containment. 
“III-B-l-c. Experiments involving the 
introduction of recombinant DNA into 
CDC Class 4** or Class 5 agents can be 
carried out at P4 containment. A USDA 
permit is required for work with Class 5 
agents***. 
“lII-B-2. Experiments in Which DNA 
from CDC Class 2 or Class 3 Agents * is 
Cloned in Nonpathogenic Prokaryotic or 
Lower Eukaryotic Host- Vector Systems. 
“III-B-2-a. Recombinant DNA 
experiments in which DNA from CDC 
Class 2 or Class 3 agents* 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 
w'ill be exempt from the Guidelines (See 
Sections IIl-D-4 and IIl-D-5). 
Experiments involving the formation of 
recombinant DNAs for certain toxin 
genes requires RAC review and NIH 
approval (see Section III-A-1), or must 
be carried out under NIH specified 
conditions as described in Appendix G. 
“lII-B-2-b. Recombinant DNA 
experiments in which DNA from CDC 
Class 4** or Class 5 agents is 
transferred into nonpathogenic 
prokaryotes or lower eukaryotes can be 
performed at P2 containment 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***. 
“III-B-3. Experiments Involving the 
Use of Whole Animal or Plant Viruses 
or Defective Animal or Plant Viruses in 
the Presence of Helper Virus in Tissue 
Culture Systems. 
“III-B-3-a. Experiments involving the 
use of whole CDC Class 2 animal 
viruses*, 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 whole CDC Class 3 animal 
viruses*, or defective CDC Class 3 
animal viruses* in the presence of 
helper virus, can be carried out at P3 
containment. 
“II1-B-3-C. Experiments involving the 
use of whole CDC Class 4 or Class 5 
animal viruses*, or defective CDC Class 
4 or Class 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***. 
“UI-B-3-d. Experiments involving the 
use of whole 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 
containment. 
“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). 
‘‘III-B-4-b. For all experiments 
involving whole animals and plants and 
not covered by III-B-4-a, the 
appropriate containment will be 
determined by the IBC. 
“IIl-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). 
“IIl-C. Experiments that Require IBC 
Notice Simultaneously with Initiation of 
Experiments. 
“For experiments in this category, a 
registration document as described in 
Section IIl-B must be dated and signed 
by the investigator and filed with the 
local IBC. The IBC shall review all such 
proposals, but IBC review prior to 
initiation of the experiment is not 
required. All experiments not included 
in categories III-A, III-B, III^, and 
subsections of these categories are to be 
considered as in category III-C. All such 
experiments can be carried out at Pi 
containment. Some explicit cases are 
listed below. 
“III-C-1. Experiments Involving Non- 
Pathogenic Prokaryotes or Non- 
Pa thogenic Lower Eukaryotic Host- 
Vector Systems. 
“Experiments involving non- 
pathogenic prokaryotes or non- 
pathogenic lower eukaryotes can be 
carried out at Pi containment. 
“III-C-2. Experiprents 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. 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 
molecules are exempt from these 
Guidelines and no registration with the 
IBC is necessary. 
“III-D-1. Those that are not in 
organisms or viruses. 
“III-D-2. Those that consist entirely of 
DNA segments from a single non- 
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