Federal Register / Vol. 44, No. 232 / Friday, November 30, 1979 / Notices 
69223 
measures provide an entirely adequate 
degree of containment. They are similar 
to those required in many countries for 
licensed handling of “exotic” plant 
viruses. 
The CaMV strain used as a cloning 
vector shall be a mutant that lacks the 
aphid transmission factor. 
The viruses or their DNA may also be 
useful as vectors to introduce genes into 
plant protoplasts. The fragility of plant 
protoplasts combined with the 
properties of the viruses provides 
' CBC - case by case (45). 
**See text 
III— C — 4. Plant Host- Vector Systems 
Other than Viruses. 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 P2 physical 
containment. The development of host- 
vector systems that exhibit a high level 
of biological containment, such as those 
using protoplasts or undifferentiated 
cells in culture, permit [2A] a decrease 
in the physical containment to Pi. 
Intact plants or propagative plant 
parts which cannot be grown in a 
standard P2 laboratory because of their 
adequate safety. Since no risk to the 
environment from the use of the DNA 
plant virus/protoplast system is 
envisaged, no special containment is 
necessary, except as described in the 
following paragraph. 
Experiments involving the use of plant 
virus 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-(3Hc).) 
large size may be grown under the Pi 
conditions described above in Section 
III— C— 3, except that (i) sterilization of 
run-off water is required where this is a 
plausible route for secondary infection 
and (ii) the standard P2 practices are 
adopted for microbiological work. 
Ill— C— 5. Fungal or Similar Lower 
Eukaryotic Host- Vector Systems. 
Certain certified HVl and HV2 host- 
vector systems appear in Appendix D. 
The containment levels for these 
systems are given in the subsections of 
Section III-A. Other systems in the 
future may be certified as HVl and HV2. 
At the time of certification, they may be 
added to Appendix D (and thus the 
containment levels for their use will be 
those of the subsections of Section III— 
A). Alternatively, at the time of their 
certification, another classification of 
containment levels for experiments 
using them may be assigned by NIH. 
In addition to the experiments 
described above, the following 
experiments may be carried out without 
the eukaryotic host (Host C) having 
been approved as an HVl host: DNA 
from Host C may be inserted into 
lambdoid phage vector or into a vector 
from a certified EK2 host-vector system 
and propagated in E. coli K-12 under Pi 
conditions. Subsequently, this 
recombinant DNA may be returned to 
Host C and propagated there under PI 
conditions. [43] 
Containment levels for other classes 
of experiments involving non-HVl 
systems may be expressly approved by 
the Director, NIH. (See Sections IV-E-1- 
b-(lHb). IV-E-l-b-(2)-{c). and IV-E-1- 
b— (3)— (b).) 
Ill— C— 6. Return of DNA Segments to a 
Higher Eukaryotic Host of Origin. DNA 
from a higher eukaryote (Host D) may 
be inserted into a lambdoid phage 
vector or into a vector from a certified 
EK2 host-vector system and propagated 
in E. coli K-12 under Pi containment 
conditions. Subsequently, this 
recombinant DNA may be returned to 
Host D and propagaged under 
conditions of physical containment 
comparable to PI and appropriate to the 
organism under study. [2A] 
III— C— 7. Transfer of cloned DNA 
Segments to Eukaryotic Organisms 
III-C-7-a. Transfer to Non-human 
Vertebrates. DNA from any 
nonprohibited source [Section I-D], 
except for greater than one quarter of a 
eukaryotic viral genome, which has 
been cloned and propagated in E. coli 
under Pi conditions, may be transferred 
with the E. coli vector used for cloning 
to any eukaryotic cells in culture or to 
any non-human vertebrate organism and 
propagated under conditions of physical 
containment comparable to Pi and 
appropriate to the organism under study 
[2A], Transfers to any other host will be 
considered by the RAC on a case-by- 
case basis [45]. 
III-C-7-b. Transfer to Higher Plants. 
DNA from any nonprohibited source 
[Section I-D] which has been cloned 
and propagated in E. coli under Pi 
conditions, may be transferred with the 
E. coli vector used for cloning to any 
higher plant organisms (Angiosperms 
and Gymnosperms) and propagated 
Table IV— Recommended Containment tor Recombinant DNA Research Using Eukaryotic Viral Vectors 
(See text for lull details) 
vector DNA 
Productive virus-cefl interactions 
Non- 
productive 
vuus-cetl 
inter- 
actions 
(46) 
Type o i DNA insert 
Prokaryotic 
Shotgun Purified 
Eukaryotic 
Shotgun 
Punned 
(47] 
Eukaryotic 
waf 
Natural 
host 
Other 
1 Potyom* 
P2 
P2 
P2 
P3 
P2 
CBC* 
P2 
Deleted Genome ...... 
P2 . 
P2 
P2 
P2 . 
P2 
CBC* 
P2 
2 SV40 
P2 
Deleted Genome 
P2 
P2 
P2 
P3 
P3 
CBC* 
P2 
3 Human Ad2 ♦ Ad5 Dieted Genome 
P3 
P3 
P3 
P3 
P3 
CSC* ... 
P2 
4 Mouse Adenovirus: (Strain FI) 
Intact Genome 
CBC* ... 
CBC* _. 
CBC* ._ 
CBC* — 
CBC* 
CBC* _ 
P2 
Deleted Ganome 
P2 
P2 
P2 
P2 
P2 
CBC* . . 
P2 
5 Insect Viruses . . 
CBC* 
CBC* .... 
C8C* 
CBC* 
CBC* 
CBC* 
i 
a 
I 
i 
« 
(**) 
(••) 
(••) 
(*•)... 
I**). .. . 
CBC* ,.. 
7 All otner potential Viral Vectors 
C8C* 
CBC* 
CBC* 
CBC* 
CBC* 
CBC* 
CBC* 
[2 09 ] 
