Federal Register / Vol. 59, No. 127 / Tuesday. July 5, 1994 / Notices 
34501 
the germ-line (transgenic animals) and 
e.xperiments involving viable 
recombinant DNA-modified 
microorganisms tested on whole 
animals. For the latter, other than 
viruses which are only vertically 
transmitted, the experiments may not be 
conducted at BLl-N containment. A 
minimum containment of BL2 or BL2- 
N is required. 
Caution — Special care should be used 
in the evaluation of containment 
conditions for some ex'perimcnts with 
transgenic animals. For example, such 
experiments might lead to the creation 
of novel mechanisms or increased 
transmission of a recombinant pathogen 
or production of undesirable traits in 
the host animal. In such cases, serious 
consideration should be given to 
increasing the containment conditions. 
Section Ill-C— 4-a. Recombinant DNA, 
or DNA or Rf4A molecules derived 
therefrom, from any source except for 
greater than two-thirds of eukaryotic 
viral genome may be transferred to any 
non-human vertebrate or any 
invertebrate organism and propagated 
under conditions of physical 
contaimnent comparable to BLl or BLl- 
N and appropriate to the organism 
under study (see Section V-B). Animals 
that contain sequences from viral 
vectors, which do not lead to 
transmissible infection either directly or 
indirectly as a result of 
complementation or recombination in 
animals, may be propagated under 
conditions of physical containment 
comparable to BLl or BLl-N and 
appropriate to the organism under 
study. Experiments involving the 
Intrc^uction of other sequences from 
eukaryotic viral genomes into animals 
are covered under Section UI-C— 4-b. For 
experiments involving recombineint 
DNA-modified Class 2, 3, 4, or 5 
organisms, see Section V-A. It is 
important that the investigator 
demonstrate that the fraction of the viral 
genome being utilized does not lead to 
productive infection. A U.S. Department 
of Agriculture permit is required for 
work with Class 5 agents (see Section V- 
R and V-T). 
Section IlI-C—4-b. For experiments 
involving recombinant DNA, or DNA or 
RNA derived therefrom, involving 
whole animals, including transgenic 
animals, and not covered by Sections 
III-C-1 or III-C—4-a, the appropriate 
containment shall be determined by the 
Institutional Biosafety Committee. 
Section III C-5. Experiments Involving 
Whole Plants 
Experiments to genetically engineer 
plants by recombinant DNA methods, to 
use such plants for other experimental 
purposes (e.g., response to stress), to 
propagate such plants, or to use plants 
together with microorganisms or insects 
containing recombinant DNA, may be 
conducted under the containment 
conditions described in Sections III-C- 
5-a through Ill-C-5-e. If experiments 
involving whole plants are not 
de.scribed in Section IIl-C-5 and do not 
fall under Sections Ill-A, in-B, or Ill-E, 
they are included in Section IIl-D. 
Note: For recombinant DNA experiments 
falling under Sections lII-C-5-a through 111- 
C-5-d, physical containment requirements 
may be reduced to the next lower level by 
appropriate biological containment practices, 
such as conducting experiments on a virus 
with an obligate insect vector in the absence 
of that vector or using a genetically 
attenuated strain. 
Section III-C-5-a. BL3-P (Plants) or 
BL2-P + biological containment is 
recommended for experiments 
involving most exotic (see Section V-W) 
infectious agents with recognized 
potential for serious detrimental impact 
on managed or natural ecosystems when 
recombinant DNA techniques are 
associated with whole plants. 
Section IIl-C-5-b. BL3-P or BL2-P 
biological containment is recommended 
for experiments involving plants 
containing cloned genomes of readily 
transmissible exotic (see Section V-W) 
infectious agents with recognized 
potential for serious detrimental effects 
on managed or natural ecosystems in 
which there exists the possibility of 
reconstituting the complete and 
functional genome of the infectious 
agent by genomic complementation in 
planta. 
Section III-C-5-c. BL4-P containment 
is recommended for experiments with a 
small number of readily transmissible 
exotic (see Section V-W) infectious 
agents, such as the soybean rust fungus 
[Phakospora pachyrhizi] and maize 
streak or other viruses in the presence 
of their specific arthropod vectors, that 
have the potential of being serious 
pathogens of major U.S. crops. 
Section III-C-5-d. BL3-P containment 
is recommended for experiments 
involving sequences encoding potent 
vertebrate toxins introduced into plants 
or associated organisms. Recombinant 
DNA containing genes for the 
biosynthesis of toxin molecules lethal 
for vertebrates at an LDjo of <100 
nanograms per kilogram body weight 
fall under Section ifi-B-l and require 
NIH/ORDA and Institutional Biosafety 
Committee approval before initiation. 
Section IIl-C-5-e. BL3-P or BL2-P + 
biological containment is recommended 
for experiments with microbial 
pathogens of insects or small animals 
associated with plants if the 
recombinant DNA-modified organism 
has a recognized potential for serious 
detrimental impact on managed or 
natural ecosystems. 
Section lll-C-6. Experiments Involving 
More than 10 Liters of Culture 
The appropriate containment will be 
decided by the Institutional Biosafety 
Committee. Where appropriate. 
Appendix K, Physical Containment for 
L^e Scale Uses of Organisms 
Containing Recombinant DNA 
Molecules, shall be used. Appendix K 
describes containment conditions Good 
Large Scale Practice through BL3-Largc 
Scale. 
Section III-C-7. Human Gene Transfer 
Experiments Not Covered bv Sections 
III-A-2. III-B~2. lII-B-3, and Not 
Considered Exempt Under Section V-U 
Certain experiments involving the 
transfer of recombinant DNA or DNA or 
RNA derived from recombinant DNA 
into one or more human subjects that 
are not covered by Sections IlI-A-2, lU- 
B-2, IU-B-3, and that are not considered 
exempt under Section V-U must be 
registered with NIH/ORDA. The 
relevant Institutional Biosafety 
Committee and Institutional Review 
Board must review and approve all 
experiments in this category prior to 
their initiation. 
Section III-D. Experiments that Require 
Institutional Biosafety Committee Notice 
Simultaneous With Initiation 
Experiments not included in Sections 
Ill-A, III-B, III-C, III-E, and their 
.subsections are considered in Section 
IIl-D. All such experiments may be 
conducted at BLl contaimnent. For 
experiments in this category, a 
registration document (see Section lIl-C) 
shall be dated and signed by the 
investigator and filed with the local 
Institutional Biosafety Committee at the 
time the experiment is initiated. The 
Institutional Biosafety Committee 
reviews and approves all such 
proposals, but Institutional Biosafety 
Committee review and approval prior to 
initiation of the experiment is not 
required (see Section FV-A). For 
example, experiments in which all 
components derived from non- 
pathogenlc prokaryotes and non- 
pathogenic lower eukaryotes fall under 
Section III-D and may be conducted ai 
BLl containment. 
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Recombinant DNA Research, Volume 19 
