Federal Register / Vol. 46. No. 210 / Friday, October 30, 1981 / Notices 
53983 
Beta carrying the structural gene for 
diphdieria toxin. 
In a letter dated July II, 1981, Dr. 
Murphy requested clarification on how 
the high containment facility located at 
Frederick. Mayland was to be used, 
j Could he be permitted to carry out the 
: experiments within the P4 facility using 
laboratory practices, equipment, and 
containment conditions specified by the 
I NIH Biosafety Committee (IBC), which 
j did not necessarily involve fuH P4 
I containment? Or. Murphy's request was 
. published for comment in the August 4, 
j 1961 Federal Register (46 FR 39771). No 
comments were received on the request 
The RAC discussed the issue at its 
September 10-11, 1981 meeting. During 
I the discussion, it was noted that 
Building 550 at the Frederick Cancer 
Resear^ Center possesses full P4 
containment barriers. The RAC judged 
that it would be appropriate for the local 
IBC to specify containment conditions 
within the facility, rather than requiring 
P4 contaiiunent for all phases of the 
work. The experimenta however, must 
be confined to DCRC Building 550. By a 
vote of eleven in favor, none opposed, 
with three abstentions, RAC 
recommended that the IBC be permitted 
to specify containment conditions for 
Dr. Murphy's experiments within 
Building 550 at the Frederick Cancer 
Resear^ Center. 
I accept this recommendation. Item 30 
in Appendix E is modified as follows: 
"30. Permission is granted to clone in 
E. coli K-12. in high containment 
building 550 at the Frederick Cancer 
Research Center, restriction fragments 
of Corynephage Beta carrying the 
structural gene for diphtheria toxin. 
Laboratory practices and containment 
equipment are to be specified by the 
IBC." 
n. Summary of Actions Under The 
Guidelines 
A. Modification of Section III-0-2 of the 
Guidelines 
Section HI-0-2 of the Guidelines is 
amended to read as follows: 
"111-0-2. Experiments Involving 
Nonpathogenic Prokaryotic and Lower 
Eukaryotic Host-Vector Systems. DNA 
from emy species nonpathogenic for 
man. animals, or plants may be cloned 
into lower eukaryotes nonpathogenic for 
man. animals, or plants at the P3 level of 
containment [2A]. DNA from any 
species nonpathogenic for man, animals, 
or plants may be cloned in o 
prokaryotes nonpathogen 'c for ntan. 
animals, or plants at the P2 level of 
containment [2Aj. Data supporting the 
contention that the donor a \d recipient 
are nonpathogenic must be ubmitted 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))." 
B. Modification of Section III-C-2-a and 
Addition of New Section III-C-7-c 
Section IIl-C-2-a is amended to read 
as follows: 
“III-C-2. Invertebrate Host-Vector 
Systems. 
"III-C-2-a. Invertebrate Viral Vectors. 
Experiments Involving invertebrate 
virus vectors can be done as follows: 
"Ul-C-2-a(l). Recombinaiit DNA 
molecules containing no more than two- 
thirds of the genome of any invertebrate 
virus [all viruses from a single Family 
(36) being considered identical (50)] may 
be propagated and maintained in cells in 
tissue culture using Pi containment. For 
such experiments, it must be shown that 
the cells lack helper virus for the 
specific Families of defective viruses 
being used. The DNA may contain 
fragments of the genomes of viruses 
fi-om more than one Family but each 
fragment must be less than two-thirds of 
a genome. 
"IIl-C-2-a-(2). Recombinants with 
less than two-thirds of the genome of 
any invertebrate virus may be rescued 
with helper virus using P2 containment 
unless it is classified by the CDC as a 
class 3 agent (1) in which case P3 
containment is required. 
"IU-C-2-a-{3). 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 
rV-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).)" 
A new Section IU-G-7-c is added as 
follows: 
"IlI-C-7-c. Transfer to Invertebrates. 
DNA fi’om any non-prohibited source 
[Section I-D], except for greater than 
one quarter of a eukaryotic viral 
genome, which has been cloned and 
propagated in E. coli K-12, may be 
transferred with the E. coli vector used 
for cloning to any eukaryotic cells in 
culture or to any invertebrate 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).” 
C. Amendment of Section I-D-6 of the 
Guidelines 
Section I-D-6 is amended to read: 
“I-D-6. Large-scale experiments [e.g., 
more than 10 liters of culture] with 
organisms containing recombinant 
DNAs other than those listed in 
Appendix C, Sections 2, 3, and 4 of the 
Guidelines, unless the recombinant 
DNAs are rigorously characterized and 
the absence of harmful sequences 
established (3). [See Section IV-E-l-b- 
(3Hd).]" 
Sections 2, 3 and 4 of Appendix C are 
amended to read as follows: 
"2. Experiments Involving E. coli K-12 
host-vector systems. Experiments which 
use E. coli K-12 host-vector systems, 
with the exception of those experiments 
listed below, are exempt from these 
Guidelines provided that (a) the E. coli 
host shall not contain conjugation 
proficient plasmids or generalized 
transducing phages, and^b) lambda or 
lambdoid or Ff bacterio-phages or 
nonconjugative plasmids [49] shall be 
ased as vectors. However, experiments 
involving the insertion Into E. coli K-12 
of DNA from prokeu^otes that exchange 
genetic Information [35] with E. coli may 
be performed with any E. coli K-12 
vector [e.g., conjugative plasmid]. When 
a nonconjugative vector is used, the E 
coli K-12 host may contain conjugation- 
proficient plasmids either autonomous 
or integrated, or generalized transducing 
phages. 
“For these exempt experiments. Pi 
physical containment conditions are 
recommended. 
"Exceptions: 
"Experiments described in Section I- 
D-1 to I-D-5 as being prohibited. 
"Experiments involving DNA from 
Class 3 organisms [1] or fiom cells 
known to be infected with these agents 
may be conducted at P3 containment. 
Lower containment levels may be 
specified by NIH. [See Section IV-E-1- 
b^2)-(e).] Experiments in this category 
require prior BBC review and approval. 
“Experiments which Increase the 
virulence and host range of a plant 
pathogen beyond that which occurs by 
natural genetic exchange. (See Section 
m-0-1.) 
“Large-scale experiments (e.g., more 
than 10 liters of culture) require prior 
IBC review and approval. 
“Experiments involving the deliberate 
cloning of genes coding for the 
biosynthesis of toxins potent for 
vertebrates. (See Appendix G.) 
[253] 
