clear idea of the location within the 
molecule of: 
(a) The sites at which DNA synthe- 
sis originates and terminates. 
(b) The sites that are cleaved by re- 
striction endonucleases. 
(c) The template regions for the 
major gene products. 
If possible the helper virus genome 
should: 
(1) Be integrated into the genome of 
a stable line of host cells (a situation 
that would effectively limit the 
growth of the vector recombinant to 
such cell lines), or 
(ii) Consist of a defective genome, or 
an appropriate conditional lethal 
mutant virus, making vector and 
helper dependent upon each other for 
propagation. 
However, neither of these stipula- 
tions is a requirement. 
(1) Polyoma virus. 
(a) Productive virus-cell interac- 
tions. 
1. Defective or Intact polyoma virus 
genomes, with appropriate helper, if 
necessary, can be used in P2 conditions 
to propagate DNA sequences from: 
(a) Bacteria of class 1 or class 2 (see 
appendix B), or their phages or plas- 
mids, except for species of bacteria 
that produce potent polypeptide 
toxins. 
(6) Prom mice. 
(c) From other eukaryotic organisms 
that do not produce potent polypep- 
tide toxins, provided the DNA segment 
is purified. 
2. Defective or intact virus genomes 
with appropriate helper, if necessary, 
can be used in P3 conditions for shot- 
gun experiments to propagate DNA se- 
quences from eukaryotic organisms, 
provided the DNA is obtained from 
uninfected cells such as embryonic or 
tissue culture cells. 
3. Experiments involving the use of 
defective polyoma virus genomes to 
propagate DNA sequences from eukar- 
yotic viruses will be evaluated by the 
Recombinant DNA Molecule Program 
Advisory Committee on a case-by-case 
basis and will be conducted under 
physical containment conditions rec- 
ommended by that committee. 
(b) Non-productive virus-cell inter- 
actions.— Defective or intact polyoma 
virus genomes can be used as vectors 
in P2 conditions to transform nonper- 
missive cells in culture. 
(2) Simian virus 40. 
(a) Productive virus-cell interac- 
tions. 
1. SV40 DNA, rendered uncondition- 
ally defective by a deletion in an es- 
sential gene, with appropriate helper, 
if necessary, can be used in P2 condi- 
tions to propagate DNA sequences 
from: 
(a) Bacteria of class 1 or class 2 <see 
appendix B). or their phages or plas- 
mids, except for species of bacteria 
NOTICES 
that produce potent polypeptide 
toxins. 
(6) Uninfected African green 
monkey kidney cells. 
2. SV40 DNA, rendered uncondition- 
ally defective by a deletion in an es- 
sential gene, with an appropriate 
helper, if necessary, can be used in P3 
conditions to propagate DNA se- 
quences from eukaryotic organisms 
(shotgun experiments or purified 
DNA) provided the DNA is obtained 
from uninfected cells such as embry- 
onic or tissue culture cells. 
3. Experiments involving the use of 
defective SV40 genomes to propagate 
DNA sequences from eukaryotic vir- 
uses will be evaluated by the Recom- 
binant DNA Molecular Program Advi- 
sory Committee on a case-by-case basis 
and will be conducted under physical 
containment conditions recommended 
by that committee. 
(b) Non-productive virus-ceU inter- 
actions.— Defective or intact SV40 gen- 
omes can be used as vectors in P2 con- 
ditions to transform nonpermissive 
cells in culture. 
(3) Human adenoviruses 2 and 5. 
(a) Productive virus-cell interac- 
tions. 
1. Human adenoviruses 2 and 5, ren- 
dered unconditionally defective by de- 
letion of at least 2 capsid genes, with 
appropriate helpers), if necessary, can 
be used in P3 conditions to propagate 
DNA sequences from: 
(a) Bacteria of class 1 or class 2 (see 
appendix B) or their phages or plas- 
mids except for species of bacteria 
that produce potent polypeptide 
toxins. 
(6) Eukaryotic organisms (shotgun 
experiments or purified DNA) pro- 
vided the DNA Is obtained from unin- 
fected cells such as embryonic or 
tissue culture cells. 
2. Experiments involving the use of 
unconditionally defective human Ad 2 
and 5 genomes to propagate DNA se- 
quences from eukaryotic viruses will 
be evaluated by the Recombinant 
DNA Molecule Program Advisory 
Committee on a case-by-case basis and 
will be conducted under physical con- 
tainment conditions recommended by 
that committee. 
(b) Non-productive virus-cell inter- 
actions.— Detective or intact human Ad 
2 and S genomes can be used as vectors 
in P2 conditions to transform non-per- 
missive cells in culture. 
(4) Murine adenovirus strain FL. 
(a) Productive virus-ceU interac- 
tions. 
1. Unconditionally defective murine 
adenoyirus strain. FL genomes, with 
appropriate helper, if necessary, can 
be used in P2 conditions to propagate 
DNA sequences from: 
(a) Bacteria of class 1 or class 2 (see 
appendix B) or their phages or plas- 
33173 
mids except for species of bacteria 
potent polypeptide toxins. 
(6) Eukaryotic organisms (shotgun 
experiments or purified DNA) pro- 
vided the DNA is obtained from unin- 
fected cells such as embryonic or 
tissue culture cells. 
2. Experiments involving the use of 
intact murine adenovirus strain FL 
genomes to propagate DNA sequences 
from prokaryotic or eukaryotic organ- 
isms will be evaluated by the Recom- 
binant DNA Molecule Program Advi- 
sory Committee on a case-by-case basis 
and will be conducted under physical 
containment conditions recommended 
by that committee. 
3. Experiments involving the use of 
unconditionally defective murine 
adenovirus strain PL genomes to prop- 
agate DNA sequences from eukaryotic 
viruses will be evaluated by the Re- 
combinant DNA Molecule Program 
Advisory Committee on a case-by-case 
basis and will be conducted under 
physical containment conditions rec- 
ommended by that committee. 
(b) Non-productive virus-ceU inter- 
actions.— Defective or Intact murine 
adenovirus strain FL genomes can be 
used as vectors in P2 conditions to 
transform non-permlssive cells in cul- 
ture. 
(5) AU. other potential viral vectors. 
(a) Experiments involving the use of 
viral DNA vectors consisting of 25 per- 
cent or less of the viral genome shall 
be used: 
1. In P2 conditions to transform non- 
permlssive cells In culture. 
2. Under physical containment con- 
ditions to be determined by the Re- 
combinant DNA Molecule Program 
Advisory Committee to propagate 
DNA sequences from prokaryotic or 
eukaryotic organisms. 
(b) Experiments involving the use of 
other intact or defective virus genomes 
to propagate DNA sequences from pro- 
karyotic or eukaryotic organisms (and 
viruses) or as vectors to transform 
non-permlssive cells will be evaluated 
by the Recombinant DNA Molecule 
Program Advisory Committee on a 
case-by-case basis and will be conduct- 
ed under physical containment condi- 
tions recommended by that commit- 
tee. 
The Recombinant DNA Molecule 
Program Advisory Committee will also 
review an experiments involving the 
use of virus vectors in animals and the 
physical containment conditions ap- 
propriate for such studies. 
b. Invertebrate host-vector systems 
in which insect viruses are used to 
propagate other DNA segments.— As 
soon u. information concerning the 
nature of the host range, infectivity, 
persistence and integration in verte- 
brate and invertebrate ceils become* 
available, experiments involving the 
use of baculo-viruses to propagate 
FEDERAL REGISTER, VOL 43, NO. 146— FRIDAY, JULY 23, 1973 
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