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Federal Register / Vol. 47, No. 102 / Wednesday. May 26. 1982 / Notices 
rvcombinant DNA molecules, if the Director. 
NIH with advice of the RAC. after 
appropriate notice and opportunity for public 
comment, finds that they do not present a 
significant risk to health or the envrionment 
(See Section rV-C-l-lHiHclk) Certain 
classes are exempt as of publication of these 
Resived Guidelines.” 
The following classes of experiments are 
exempt under Section Ul-D-5 of the 
Guidelines: 
Appendix C-L Recombinant DNAs in 
Titaue Culture. Recombinant BNA molecules 
derived entirely horn non-viral components 
(that is. no component is derived from a 
eukaryotic virus), that are propagated and 
maintained in cells in tissue oJture are 
exempt from these Guidelines with the 
exceptions listed below. 
Exceptions. Experiments described in 
Section Ul-A which require specific RAC 
review and NIH approval before initiation of 
the experiment. 
Experiments involving DNA from Class 3. 
4. or S organisms (1) or cells known to be 
infected with these agents. 
Experiments involving the deliberate 
introduction of genes coding for the 
biosynthesis of molecules toxic for 
vertebrates. (See Appendix F.) 
Appendix C-U. Experiments Involving E 
coli K-12 Host- Vector Systems. Experiments 
which use £ coH K-12 host-vector systems, 
with the exception of thoee experiments 
listed below, are exempt from these 
Guidelines provided that (a) the £ coli hoet 
shall not contain conjugation proficient 
plasmids or generalized transdudog phages, 
end (b) lambda or lambdoid or Pf bacterio- 
phages or nooconjugaJve plasmids |2| shall 
be used as vectors. However, experiments 
Involving the insertion into £ coli K-12 of 
DNA from piokaryotes that exchange genetic 
information (3] with £ coU may be 
performed with any £ coli K-12 vector (e.g.. 
conjugativa plasmid). When a nonconjugative 
vector is us^ the £ coli K-12 host may 
contain conjugation-proficient plasmids ether 
autonomous or integrated, or generalized 
transducing phages. 
For these exempt experiments, PI physical 
containment conditions are recommended. 
Exceptions. Experiments described in 
Section m-A which require a specific RAC 
review and NIH approval before Initiation of 
the experiment. 
Exp^ments involving DNA from class 3, 4. 
or 5 organisms (1] or from cells known to be 
infected with these agents may be conducted 
under containment conditions specified in 
Section Ul-B-2 with prior IBC review and 
approval 
Large-scale experiments (e.g.. more than 10 
liters of culture) require prior IBC review and 
approval. (See Section in-B-5.) 
Experiments involving the deliberate 
cloning of genes coding for the the 
biosynthesis of molecules toxic for 
vertebrates. (See Appendix F.) 
Appendix C-IU. Experiments Involving 
Saccharomyces cerevisiae Host- Vector 
Systems. Experiments which use 
^ccbaromyces cerevisiae host-vector 
systems, with the exception of experiments 
listed below, are exempt from these 
Guidelines provided that laboratory strains 
are used. 
For these exempt experiments. PI physical 
containment conditions are recommended. 
Exemptions. Experiments described in 
Section Ill-A which require speciBc RAC 
review and NIH approval before initiation of 
the experiment 
Experiments involving Gass 3. 4 or 5 
organisms [1] or cells known to be infected 
with these agents may be conducted under 
containment conditions specified in Section 
IU-B-2 with prior IBC review and approval. 
Large-scale experiments (e g., more than 10 
liters of culture) require prior IBC review and 
approval. (See Section Dl-B-S.) 
Experiments involving the deliberate 
cloning of genes coding for the biosynthesis 
of molecules toxic for vertebrates. (See 
Appendix F.) 
Appendix C-TV. Experiments Involving 
Bacillus subtilis Host- Vector Systems. Any 
asporogenic Bacillus subtilis strain which 
does not revert to a sporeformer with a 
frequency greater than 10* * can be used for 
cloning DNA. with the exception of those 
experiments listed below. Indigenous 
Bacillus plasmids and phages, whose host- 
range does not include Bacillus cereus or 
Bacillus anthracis, may be used as vectors. 
For these exempt experiments PI physical 
contaiment conditions are recommended. 
Exceptions. Experiments described in 
Section Ill-A which require specific RAC 
review and approval before initiation of the 
experiment. 
Experiments involving Class 3. 4. or 5 
organisms [1] or cells known to be infected 
with these agents may be conducted under 
contaiiunent conditions specified by Section 
in-B-2 with prior IBC review and approval. 
Large-scale experiments (eg., more than 10 
liters of culture) require prior IBC review and 
approval (See Section IlI-B-5.) 
Experiments involving the deliberate 
cloning of genes coding for the biosynthesis 
of molecules toxic for vertebrates. (See 
Appendix F.) 
Appendix C- V. Footnotes and References of 
Appendix C 
1. The original reference to organisms as 
Class 1. 2. 3. 4. or 5 refers to the calssification 
in the publication Classification of Etiologic 
Agents on the Basis of Hazard. 4th Edition. 
)uly 1974: U.S. Department of Health. 
Education, and Welfare. Public Health 
Service. Centers for Disease Control OfTice 
of Biosafety. Atlanta. Georgia 30333. 
The Director. NIH. with advice of the 
Recombinant DNA Advisory Committee, may 
revise the classirication for the purposes of 
these Guidelines (see Section rV-C-l-b-(2)- 
(d)). The revised list of organisms in each 
class is reprinted in Appendix B to these 
Guidelines. 
2. subset of non-conjugative plasmid vectors 
are also poorly mobilizable (e.g.. pBR322. 
pBR313). Where practical these vectors 
should be employed. 
3. Defined as observable under optimal 
laboratory conditions by transformation, 
transduction, phage infection, and/or 
conjugation with transfer of phage, plasmid, 
and/or chromosomal genetic information. 
Note that this definition of exchange may be 
less stringent than that applied to exempt 
organisms unser Section 111-0-4. 
Apendix D — Actions Taken Under the 
guidelines 
As noted in the subsections of Section IV- 
C-l-b-(l). the Director. NIH, may take 
certain actions with regard to the Guidelines 
after the issues have been considered by the 
RAG 
Some of the actions taken to date include 
the following: 
Appendix D-I. Permission is granted to 
clone Foot-and-Mouth disease virus in the 
EKl host-vector system consisting of E. coli 
K-12 and the vector pBR322, all work to be 
done at the Plum Island Animal disease 
Center. 
Appendix D-IL Certain specified clones 
derived from segments of the Foot-and-Mouth 
disease Virus may be transferred from Plum 
Island Animal disease Center to the facilities 
of Genentech, Inc., of South San Francisco, 
California. Further development of the clones 
at Genetech has been approved under PI -t- 
EKl conditions. 
Appendix D-ill. The Rd strain of 
hemophilus influenzae can be used as a host 
for the propagtin of the cloned Tn 10 tet R 
gene derived from £ coli K-12 employing the 
non-conjugative Haemophilus plasmid 
pRSF0885. under Pi conditions. 
Appendix D-IV. Permission is granted to 
clone certain subgenomic segments of Foot- 
and-Mouth Disease Virus in HVl Bacillus 
subtilis and Saccharomyces cerevisiae host- 
vector systems under Pi conditions at 
Genetech. Inc., South San Francisco, 
California. 
Appendix D-V. Permission is granted to Dr. 
Ronald Davis of Stanford University to field 
test com plants modified by recombinant 
DNA teclmiques under specified containment 
conditions. 
Appendix D-VI. Permission is granted to 
clone in £ coli K-12. under PI physical 
containment conditions, subgenomic 
segments of Rift Valley Fever Virus subject to 
conditions which have been set forth by the 
RAG 
Appendix D-Vn. Attenuated laboratory 
strains of Salmonella typhimurium may be 
used under Pi physical containment 
conditions to screen for the Saccharomyces 
cerevisiae pseudouridine synthetase gene. 
The plasmid YEpl3 will be employed as the 
vector. 
Appendix D-VIII. Permission is granted to 
transfer certain clones of subgenomic 
segments of Foot-and-Mouth Disease Virus 
from Plum Island Animal Disease Center to 
the laboratories of Molecular Genetics, Inc., 
Minnetonka. Minnesota, and to work with 
these clones under PI containment 
conditions. Approval is contingent upon 
review of data on infectivity testing of the 
clones by a working group of the RAC. 
Appendix E — Certified Host-Vector Systems 
While many experiments using £ coli K-12, 
Saccharomyces cerevisiae and Bacillus 
subtilis are currently exempt from the 
Guidelines under Exemption III-D-5, some 
derivatives of these host-vector systems were 
previously classified as HVl or HV2. A 
listing of those systems follows. 
HVl. The following plasmids are accepted 
as the vector components of certified B. 
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