Federal Register / Vol. 45, No. 20 / Tuesday, January 29, 1980 / Notices 
6737 
NLH 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-(3Hc).) 
Ill— C— 1— f. Non viral 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. 
IIl-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 [45) and will be 
conducted under the recommended 
physical containment conditions. (See 
Section IV-E-l-b-(3)-(c).} 
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 cells from 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 
Table IV 
evaluated by NIH on a case-by-case 
basis [45). 
Ill— 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 
molecule weight of 4.5 X 10 6 , and Bean 
Golden Mosaic Virus (BGMV) and 
related viruses with small (<10 s 
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 ta plants. For this 
reason, they are most unlikely to be 
accidentally transmitted from spillage of 
purified virus preparations. 
Recommended Containment for Recombinant ENA Research Using Eukaryotic Viral Vectors 
(See text for full details) 
Vector ENA 
Productive virus-cell interactions 
Nonproductive 
virus-cell 
interactions (46) 
'type of DNA insert 
Prokaryotic Eukaryotic 
Shotgun Purified Shotgun Purif ied[47) 
Natural host Other 
Eukaryotic 
viral 
TT 
Polyana 
Intact Genome 
P2 
P2 
P2 
P3 
P2 
CBC* 
P2 
Deleted Genome 
P2 
P2 
P2 
P2 
P2 
CBC* 
P2 
2. 
SV40 
Intact Gencme 





P2 
Deleted Genome 
P2 
P2 
P2 
P3 
P3 
CBC* 
P2 
3. 
Human M2 and Ad5 
Deleted Genome 
P3 
P3 
P3 
P3 
P3 
CBC* 
P2 
4. 
House Adenovirus 
(Strain EL) 
Intact Genome 
CBC* 
CBC* 
CBC* 
CBC* 
CBC* 
CBC* 
P2 
Deleted Genome 
P2 
P2 
P2 
P2 
P2 
CBC* 
P2 
5. 
Insect Viruses 
CBC* 
CBC* 
CBC* 
CBC* 
CBC* 
CBC* 
— 
6 . 
Plant Viruses 
(CaMV and BOW) 
** 
• * 
** 
• * 
** 
CBC* 
— 
7. 
All other potential 
Viral vectors 
CBC* 
CBC* 
CBC* 
CBC* 
CBC* 
CBC* 
CBC* 
*CBC » case-by-case [45] 
“See text 
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- 
[ 29 ] 
