Federal Register / Vol. 46, No. 233 / Friday, December 4, 1981 / Notices 
59407 
containment [2A]. DNA from any 
species nonpathogenic for man, animals, 
or plants may be cloned into 
prokaryotes nonpathogenic for man, 
animals, or plants at the P2 level of 
containment [2A]. Data supporting the 
contention that the donor and recipient 
are nonpathogenic must be submitted to 
the local IBC. Lower levels of physical 
containment may be assigned by ORDA 
on a case-by-case basis for specific 
donor-recipient combinations (see 
Section IV-E-l-b-(3)-(h)). 
III-A. Classification of Experiments 
Using Certain HVl and HV2 Host- 
Vector Systems. Certain HVl and HV2 
host-vector systems are assigned 
containment levels as specified in the 
subsections of this Section III-A. Those 
so classified as of publication of these 
revised Guidelines are listed in 
Appendix D. An updated list may be 
obtained from the Office of 
Recombinant DNA Activities, National 
Institutes of Health, Bethesda, Maryland 
20205. 
III-A-1. Shotgun Experiments. These 
experiments involve the production of 
recombinant DNAs between the vector 
and portions of the specified cellular 
source, preferably a partially purified 
fraction. Care should be taken either to 
preclude or eliminate contaminating 
microorganisms before isolating the 
DNA. 
III-A-l-a . Eukaryotic DNA 
Recombinants. 
III-A-l-a-(l). Primates. P2 physical 
containment -I- an HV2 host-vector or 
P3-I-HV1. 
lIl-A-l-a-(2). Other Mammals. P2 
physical containment -Fan HV2 host- 
vector or P3-I-HV1. 
III-jA-l-a-(3). Birds. P2 physical 
containment -Fan HV2 host-vector, or 
P3-FHV1. 
III-A-l-a-(4). Cold-Blooded 
Vertebrates. P2 physical 
containment -Fan HVl host-vector or 
Pi -fHV 2. If the eukaryote is known to 
produce a potent polj^ieptide toxin, [34] 
the containment shall be increased to 
P3-FHV2. 
IIl-A-l-a-(5). Other Cold-Blooded 
Animals and Lower Eukaryotes. This 
large class of eukaryotes is divided into 
two groups: 
III-A-l-a-(5]-{a). Species that are 
known to produce a potent polypeptide 
toxin[34] that acts in vertebrates, or are 
known pathogens listed in Class 2,(1] or 
are known to carry such pathogens must 
use P3 physical containment -F an HV2 
host-vector. When the potent toxin is 
not a polypeptide and is likely not to be 
the product of closely linked eukaryote 
genes, containment may be reduced to 
P3-FHV1 or P2-I-HV2. Species that 
produce potent toxins that affect 
invertebrates or plants but not 
vertebrates require P2 -fHV 2 or 
P3-FHV1. Any species that has a 
demonstrated capacity for carrying 
particular pathogenic microorganisms is 
included in this group, unless the 
organisms used as the source of DNA 
have been shown not to contain those 
agents, in which case they may be 
placed in the following group.[2A] 
III-A-l-a-(5}-(b). The remainder of 
the species in this class including plant 
pathogenic or symbiotic fungi that do 
not produce potent toxins: P2-FHV1 or 
P1-FHV2. However, any insect in this 
group must be either (i) grown under 
laboratory conditions for at least 10 
generations prior to its use as a source 
of DNA, or (ii) if caught in tlie wild, must 
be shotvn to be free of disease-causing 
microorganisms or must belong to a 
species that does not carry 
microorganisms causing disease in 
vertebrates or plants.[2A] If these 
conditions caimot be met, experiments 
must be done under P3-FHV1 or 
P2-FHV2 containment. 
III-A-l-a-(6). Plants. P2 physical 
containment -Fan HVl host-vector, or 
P1-FHV2. If the plant source makes a 
potent polypeptide toxin,[34] the 
containment must be raised to P3 
physical containment -Fan HV2 host- 
vector. When the potent toxin is not a 
olypeptide and is likely not to be the 
roduct of closely linked plant genes, 
containment may be reduced to 
P3-FHV1 or P2-FHV2.[2A] 
III-A-l-b. Prokaryotic DNA 
Recombinants. P2-FHV1 or P1-FHV2 for 
experiments with phages, plasmids and 
DNA from nonpathogenic prokaryotes 
which do not produce polypeptide 
toxins.[34] P3-FHV2 for experiments 
with phages, plasmids and DNA from 
Class 2 agents.[l] 
III-A-2-a. Viruses of Eukaryotes 
(summary given in Table III; see also 
exception given at asterisk at end of 
Appendix D). 
III-A-2-a-(l). DNA Viruses. 
III-A-2-a-(l)-(a). Nontransforming 
viruses. 
III-A-2-a(l)-(a)-(l). Adeno- 
Associated Viruses, Minute Virus of 
Mice, Mouse Adenovirus (Strain FL), 
and Plant Viruses.[48] PI physical 
containment -F and HVl host- vector shall 
be used for DNA recombinants 
produced with (i) the whole viral 
genome, (ii) subgenomic DNA segments, 
or (iii) purified cDNA copies of viral 
mRNA.[37] 
III-A-2-a-(l)-(a)-(2). Hepatitis B. 
IIl-A-2-a-(l)-(a)-(2)-(o). Pi physical 
containment -Fan HVl host-vector shall 
be used for purified subgenomic DNA 
segments. [38] 
III-A-2-a-(l)-(a)-(2)-(Z>). P2 physical 
containment an HV2 host-vector, or 
P3-FHV1, shall be used for DNA 
recombinants produced with the whole 
viral genome or with subgenomic 
segments that have not been purified to 
the extent required in footnote 38. 
III-A-2-a-(l)-(a)-(2)-(c). P2 physical 
containment an HVl Host-vector shall 
be used for DNA recombinants derived 
from purified cDNA copies of viral 
mRNA.[37] 
III-A-2-a-(l)-(a)-(5). Other 
Nontransforming Members of Presently 
Classified Viral Families.[36j 
III-A-2-a-(l)-(a)-(5)-(o). Pi physical 
containment -Fan HVl host- vector shall 
be used for (i) DNA recombinants 
produced with purified subgenomic 
DNA[38] segments or (ii) purified cDNA 
copies of viral mRNA.[37] 
in-A-2-a-(l)-(a)-(5)-(i>). Pi physical 
containment -fan HVl host-vector shall 
be used for DNA recombinants 
produced with the whole viral genome 
or with subgenomic segments that have 
not been purified to the extent required 
in footnote 38. 
III-A-2-a-(l)-(b). Transforming 
Viruses. [37A] 
ni-A-2-a-(l)-(b)-(l). Herpes Saimiri, 
Herpes Ateles, and Epstein Barr 
Virus.[39] 
IlI-A-2-a-(l)-(b)-(l)-(a). Pi physical 
containment -F an HVl host-vector shall 
be used for DNA recombinants 
produced with purified nontransfprming 
subgenomic DNA segments. [38] 
III-A-2-a-(l)-(b)-(J)-(i). P2 physical 
containment -F an HVl host-vector shall 
be used for (i) DNA recombinants 
produced with purified subgenomic 
DNA segments containing an entire 
transforming gene [38] or (ii) purified 
cDNA copies of viral mRNA.[37] 
IIl-A-2-a-(l)-(b)-(7)-(c). P3 physical 
containment -f an HVl host-vector, or 
P2 -f HV2, shall be used for DNA 
recombinants produced with the whole 
viral genome or with subgenomic 
segments that have not been purified to 
the extent required in footnote 38. 
III-A-2-a-(l)-(b)-(2). Other 
Transforming Members of Presently 
Classified Viral Families.[36] 
III-A-2-a-(l)-(b)-(2)-(o). Pi physical 
containment -F an HVl host-vector shall 
be used for DNA recombinants 
produced with purified nontransforming 
subgenomic DNA segments. [38] 
IlI-A-2-a-(l)-(b)-(2)-(Z7). P2 physical 
containment -f an HVl host-vector shall 
be used for (i) DNA recombinants 
produced with the whole viral genome, 
(ii) subgenomic DNA segments 
containing an entire transforming gene, 
(iii) purified cDNA copies of viral 
mRNA, [37] or (iv) subgenomic segments 
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