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Federal Register / Vol. 44, No. 232 / Friday, November 30, 1979 / Notices 
are not derived from eukaryotic viruses. 
In the latter case, such experiments will 
be evaluated by NIH on a case-by-case 
basis[45] and will be conducted under 
the prescribed physical and biological 
containment conditions. (See Section 
I V— E— 1— b— (3)— (c).) 
III-C-1-d. Murine Adenovirus Strain 
FL. 
Ill— C— 1— d — (1). Productive Virus-Cell 
Interactions. 
Ill— C— 1— d— (1)— (a). Unconditionally 
defective murine adenovirus strain FL 
genomes, with appropriate helper, can 
be used in P2 conditions to propagate 
DNA sequences from: 
III— C— 1— d— (1)— (a)— bacteria of Class 
1 or Class 2[1] or their phages or 
plasmids except for those that produce 
potent polypeptide toxins;[34] 
III-C-l-d-(l)-(a)-(2}. eukaryotic 
organisms that do not produce potent 
polypeptide toxins[34] (shotgun 
experiments or purified DNA). 
Ill— C— 1— d— (1)— (b). Experiments 
involving the use of whole murine 
adenovirus strain FL genomes to 
propagate DNA sequences from 
prokaryotic or eukaryotic organisms will 
be evaluated by NIH on a case-by-case 
basis[45] and will be conducted under 
the prescribed physical and biological 
containment conditions. (See Section 
IV-E-l-b-(3)-(c).) 
Ill-C-l-d-l-(c). Experiments 
involving the use of unconditionally 
defective murine adenovirus strain FL 
genomes to propagate DNA sequences 
from eukaryotic viruses will be 
evaluated by NIH on a case-by-case 
basis [45] and will be conducted under 
the prescribed physical and biological 
containment conditions. (See Section 
IV-E-l-b-(3)-(c).) 
Ill— C— 1— d— (2). Nonproductive Virus- 
Cell Interactions. Defective or whole 
murine adenovirus strain FL genomes 
can be used as vectors in P2 conditions 
when production of viral particles 
cannot occur (e.g., transformation of 
nonpermissive cells or propagation of an 
unconditionally defective recombinant 
genome in the absence of helper), 
provided the inserted DNA sequences 
are not derived from eukaryotic viruses. 
In the latter case, such experiments will 
be evaluated by NIH on a case-by-case 
basis [45] and will be conducted under 
the prescribed physical and biological 
containment conditions. (See Section 
IV-E-l-b-(3)-(c).) 
III-C-1-e. All Other Potential Viral 
Vectors. 
Ill— C— 1— e— (1). Experiments involving 
recombinant DNA molecules containing 
viral DNA segments consisting of 25% or 
less of the virus genome can be done: 
III— C— 1— e— (1 )— (a) . In P2 conditions 
when the recombinant DNA is to be 
integrated into the cell genome or is 
known to replicate as a plasmid in cells 
in culture, provided the additional DNA 
sequences are not derived from a 
eukaryotic virus. In the latter case, such 
experiments will be evaluated by NIH 
on a case-by-case basis [45] and will be 
conducted under the prescribed physical 
and biological containment conditions. 
(See Section IV— E— 1— b— (3)— (c).) 
III-C-l-e-(l)-(b). Under physical and 
biological containment conditions to be 
determined by NIH [45] when a viral 
helper will be used to propagate DNA 
sequences from prokaryotic or 
eukaryotic organisms. (See Section IV- 
E— 1— b— (3)— (c).) 
III-C-l-e-(2). Experiments involving 
the use of other whole or defective virus 
genomes to propagate DNA sequences 
from prokaryotic or eukaryotic 
organisms (and viruses), or as vectors to 
transform nonpermissive cells, will be 
evaluated by NIH on a case-by-case 
basis [45] and will be conducted under 
the prescribed physical and biological 
containment conditions. (See Section 
IV-E-l-b-(3)-(c).) 
NIH will also review on a case-by- 
case basis [45] all experiments involving 
the use of virus vectors in animals and 
will prescribe the physical and 
biological containment conditions 
appropriate for such studies. (See 
Section IV-E-l-b-(3)-(c).) 
Ill— C— 1— f. Nonviral Vectors. 
Organelle, plasmid, and chromosomal 
DNAs may be used as vectors. DNA 
recombinants formed between such 
vectors and host DNA, when propagated 
only in that host (or a closely related 
strain of the same species), are exempt 
from these Guidelines (see Section I-E). 
DNA recombinants formed between 
such vectors and nonviral DNA from 
cells other than the host species require 
only Pi physical containment for cells in 
culture since vertebrate cells in tissue 
culture inherently exhibit a very high 
level of containment. Recombinants 
involving viral DNA or experiments 
which require the use of the whole 
animals will be evaluated by NIH on a 
case-by-case basis [45]. 
Ill— C— 2. Invertebrate Host-Vector 
Systems. 
III-C-2-a. Insect Viral Vectors. As 
soon as information becomes available 
on the host range restrictions and on the 
infectivity, persistence, and integration 
of the viral DNA in vertebrate and 
invertebrate cells, experiments involving 
the use of insect viruses to propagate 
DNA sequences will be evaluated by 
NIH on a case-by-case basis [45] and 
will be conducted under the 
recommended physical containment 
conditions. (See Section IV-E-l-b-(3)- 
(c)0 
Ill— C— 2— b. Nonviral Vectors. 
Organelle, plasmid, and chromosomal 
DNAs may be used as vectors. DNA 
recombinants formed between such 
vectors and host DNA, when propagated 
only in that host (or a closely related 
strain of the same species), are exempt 
from these Guidelines (see Section I-E). 
DNA recombinants formed between 
such vectors and DNA from cells other 
than the host species require Pi physical 
containment for invertebrate cells in 
culture since invertebrate cells in culture 
inherently exhibit a very high level of 
containment. Experiments which require 
the use of whole animals will be 
evaluated by NIH on a case-by-case 
basis [45], 
III— C— 3. Plant Viral Host-Vector 
Systems. The DNA Plant viruses which 
could currently serve as vectors for 
cloning genes in plants and plant cell 
protoplasts are Cauliflower Mosaic 
Virus (CaMV) and its close relatives 
[2A] which have relaxed circular 
double-stranded DNA genomes with a 
molecular weight of 4.5 X10 6 , and Bean 
Golden Mosaic Virus (BGMV) and 
related viruses with small (<10 6 
daltons) single-stranded DNA genomes. 
CaMV is spread in nature by aphids, in 
which it survives for a few hours? 
Spontaneous mutants of CaMV which 
lack a factor essential for aphid 
transmission arise frequently. BGMV is 
spread in nature by whiteflies, and 
certain other single-stranded DNA plant 
viruses are transmitted by leafhoppers. 
The DNA plant viruses have narrow 
host ranges and are relatively difficult to 
transmit mechanically to plants. For this 
reason, they are most unlikely to be 
accidentally transmitted from spillage of 
purified virus preparations. 
When these viruses are used as 
vectors in intact plants, or propagative 
plant parts, the plants shall be grown 
under Pi conditions — that is, in either a 
limited access greenhouse or plant 
growth cabinet which is insect- 
restrictive, preferably with positive air 
pressure, [2A] and in which an insect 
fumigation regime is maintained. Soil, 
plant pots, and unwanted infected 
materials shall be removed from the 
greenhouse or cabinet in sealed insect- 
proof containers and sterilized. It is not 
necessary to sterilize run-off water from 
the infected plants, as this is not a 
plausible route for secondary infection. 
When the viruses are used as vectors in 
tissue cultures or in small plants in 
axenic cultures, no special containment 
is necessary. Infected plant materials 
which have to be removed from the 
greenhouse or cabinet for further 
research shall be maintained under 
insect-restrictive conditions. These 
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