17198 
Federal Register / Vol. 47, No. 77 / Wednesday, April 21. 1902 / Notices 
PootnotM and refaraocM of Appendix B 
* A USOA permit, required for import and 
Interetate commerce of pathogens, may be 
obtained from the Animal and Plant Health 
inspection Service, USOA. Federal Building. 
HyatUville. MD 20782. 
** Since the publication of the 
clasaificatlon in 1974 |1), the Acdnomycetet 
have been reclassified as bacterial rather 
than fungal agenta. 
*** All activities, including storage of 
variola and whitepox are restricted to the 
single national facility (World Health 
Organisation (WHO) Collaborating Center 
for Smallpox Resear^ Center for Disease 
Control in Atlanta). 
1. ClattificoUon of Eliotogic Agenta on the 
Baaia of Hazard. (4th Edition. |uly 1974). U.S 
Department of Health. Education and 
Welfare. Public Health Service. Center for 
Disease Control Office of Biosafety. Atlanta. 
Georgia S0333. 
2. NaUona! Cancer Institute Safety 
Standards for Research Involving Oncogenic 
Viruses (October 1974). U.S. Department of 
Health. Question, and Welfare Publication 
No. (NIH) 75-79a 
3. U.S Department of Agriculture. Animal 
and Plant Health Inspection Service. 
1 
I Appendix C — Exemptioos Under II1-0-4 
Section UI-D-8 steles that exempt from 
these Guidelines are "Other classes of 
recombinant DNA molecules, if the Director. 
NIH. with advice of the RAC after 
' appropriate notice and opportunity for public 
I comment, finds that they do not present a 
i significant risk to health or the environment. 
I (See Section rV-E-l-b-(l)-<d)). Certain 
I classes are exempt as of publication of these 
Revised Guidelines." 
, The following classes of experiments are 
j exempt under Section iU-D-6 of the 
Guidelines: 
I 1. Recombinant DNAs in Tissue Culture. 
I Recombinant DNA molecules derived 
I I entirely from non-viral components (that is. 
I no component is derived from a eukaryotic 
' vtnisl that are propagated and maintained in 
{ ceils In tissue culture are exempt from these 
I Guidelines with the exceptions listed below. 
, Exceptions. Experiments described in 
I Section fll-A which require specific RAC 
I review and NIH approval before Initiation of 
the experiment. 
Experiments Involving DNA from Class 3. 
4, or 5 organisms (1) or cells known to be 
I infected with these agents. 
Expenments involving the deliberate 
! introdurlton of genes coding for the 
I biosynthesis of toxins potent for vertebrates, 
t (See Appendix F.) 
. 2 Expenments Involving E coli K-t2 Host- 
I Vector Systems. Expenments which use E. 
I coll K-12 host-vector systems, with the 
I exception of those experiments listed below. 
are exempt from these Guidelines provided 
I that (a) the E coli host shall not contain 
‘ conjugation proficient plasmids or 
I generalized transducing phages, and (b) 
I lambda or lambdoid or Ff bacteriophages or 
I nonconjugalive plasmids (49) shall be used as 
’ vectors However, expenments involving the 
I Insertion into £ coli K-12 of DNA from 
prokaryotes that exchange genetic 
information (35) with E. coli may by 
performed with any £ coli K-12 vector (e g., 
conjugative plasmid). When a nonconjugative 
vector is used, the £ coli K-12 host may 
contain conjugation-proficient plasmids 
either autonomous or integrated, or 
generalized transducing phages. 
For these exempt experiments. Pi physical 
containment concLtions are recommended. 
Exceptions. Experiments described in 
Section lll-A which require speciRc RAC 
review and NIH approval before initiation of 
the experiment 
Experiments 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 lll-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 IIl-B-5.) 
Experiments involving the deliberate 
cloning of genes coding for the biosynthesis 
of toxins potent for vertebrates. (See 
Appendix F.) 
3. Experiments Involving Saccharomyces 
cerevisioe Host- Vector Systems. Experiments 
which use Saccharomyces cerevisioe 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. 
Exceptions. Experiments described in 
Section lll-A which require specific RAC 
review and NIH approval before initiation of 
the experiment. 
Experiments involving CDC Class 3. 4. or 5 
organisms (1) or cells known to be infected 
with these agents may be conducted under 
containment conditions specified in Section 
lll-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 IIl-B-5.) 
Experiments Involving the deliberate 
cloning of genes coding for the biosynthesis 
of toxins potent for vertebrates. (See 
Appendix F.) 
4. 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 
containment conditions are recommended. 
Exceptions. Experiments described in 
Seebon III-A which require specific RAC 
review and approval before initiation of the 
experiment. 
Experiments involving CDC Class 3. 4. or 5 
organisms (1| or cells known to be infected 
with these agents may be conducted under 
containment conditions specified by Section 
III-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 III-B-5.) 
Experiments involving the deliberate 
cloning of genes coding for the biosynthesis 
of toxins potent for vertebrates. (See 
Appendix F.) 
Appendix D — Actions Taken Under The 
Guidelines 
As noted (n the subsections of Sections IV- 
E-l-b-(l) and IV-E-l-b-(2). the Director, 
NIH. may take certain actions with regard to 
the Guidelines after consideration by the 
RAC. 
Some of the actions taken to date include 
the following: 
1. Permission is granted tb clone Foot-and- 
Mouth Disease Virus in the EKI host-vector 
system consisting of £ coli K-12 and the 
vector pBR322. all work to be done at the 
Plum Island Animal Disease Center. 
2. 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 
Genenlech. Inc., of South San Franciso. 
California. Further development of the clones 
at Cenentoch has been approved under 
Pi -i- EKI conditions. 
3. The Rd strain of Hemophilus influenzae 
can be used as a host for the propagation of 
the cloned Tn 10 let R gene derived from £ 
coH K-12 employing the non-conjugative 
Haemophilus plasmid. pRSF0885. under Pi 
conditions. 
4. Permission is granted to clone certain 
subgenomic segments of Fool-and-Mouth 
Disease Virus in HVI Bacillus Subtilis and 
Saccharomyces cerevisioe host-vector 
systems under Pi conditions at Genenlech. 
Inc., South San Francisco. California. 
5. Permission is granted to Dr. Ronald 
Davis of Stanford University to field test com 
plants modified by recombinant DNA 
techniques under specified containment 
conditions. 
8. Permission is granted to clone in £ co/i 
K-12. under Pi physical containment 
conditions, subgenomic segments of Rift 
Valley Fever Virus subject to conditions 
which have been set forth by the RAC. 
7. Attenuated laboratory strains of 
Salmonella typhimurium may be used under 
Pi physical containment conditions to screen 
for the Saccharomyces cerevisioe 
pseudouridine synthetase gene. The plasmid 
YEpl3 will be employed as the vector. 
8. 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 cerevisioe and Bacillus 
subtilis are currently exempt from the 
Guidelines under Exemption IIl-D-5. some 
derivatives of these host-vector systems were 
previously classified as HVI or HV2. A 
listing of those systems follows: 
HVI — The following plasmids are accepted 
us the vector components of certified B. 
(429) 
