Federal Register / Vol. 48, No. 106 / Wednesday, June 1, 1983 / Notices 
24567 
biosynthesis of molecules toxic for 
vertebrates. (See Appendix F.) 
Appendix C— III. Experiments 
Involving Saccharomyces cerevisiae 
Host-Vector Systems. Experiments 
which use Saccharomyces 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. 
Exceptions 
Experiments described in Section III- 
A which require specific RAC review 
and NIH 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 containment 
conditions specified in 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 molecules toxic for 
vertebrates. (See Appendix F.) 
Appendix C-IV. Experiments 
Involving Bacillus subti/is Host-Vector 
Systems. Any asporogenic Bacillus 
subti/is 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 Bascillus 
plasmids and phages, whose host-range 
does not include Bacillus cereus or 
Bacillus antbracis. may be used as 
vectors. 
For these exempt experiments Pi 
physical containment conditions are 
recommended. 
Exceptions 
Experiments described in Section III- 
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 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 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 classification 
in the publication Classification of Etiologic 
Agents on the Basis of Hazard. 4th Edition. 
July 1974; U.S. Department of Health, 
Education, and Welfare, Public Health 
Service, Centers for Disease Control, Office 
of Biosafety. Atlanta, Ceorgia 30333. 
The Director, NIH. with advice of the 
Recombinant DNA Advisory Committee, may 
revise the classification for the purposes of 
these Guidelines (see Section IV-C-l-b-(2)- 
(d)). The revised list of organisms in each 
class is reprinted in Appendix B to these 
Guidelines. 
2. A subset of non-conjugative plasmid 
vectors sire 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 under Section III— D — 4. 
Appendix 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 RAC 
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— II. 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 Sooth San Francisco, 
California. Further development of the 
clones at Genentech has been approved 
under Pi -f EKl conditions. 
Appendix D-I II. The Rd strain of 
Hemophilus influenzae can be used as a 
host for the propagation of the cloned 
Tn 10 tet R gene derived from E. 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 Diesease Virus in HVl 
Bacillus sub til is and Saccharomyces 
cerevisiae host-vector systems under Pi 
conditions at Genentech, Inc., South San 
Francisco, California. 
Appendix D-V. Permission is granted 
to Dr. Ronald Davis of Stanford 
University to field test corn plants 
modified by recombinant DNA 
techniques under specified containment 
conditions. 
Appendix D-VL Permission is granted 
to clone in E. 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 RAC. 
Appendix D-VII. 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 D-IX. Permission is granted 
to Dr. John Sanford of Cornell University 
to field test tomato and tobacco plants 
transformed with bacterial [E. coli K-12) 
and yeast DNA using pollen as a vector. 
Appendix D-X. Permission is granted 
to Drs. Steven Lindow and Nickolas 
Panopulos of the University of 
California, Berkeley, to release under 
specified conditions Pseudomonas 
syringae pv. syringae and Erwinia 
herbicola carrying in vitro generated 
deletions of all or part of the genes 
involved in ice nucleation. 
Appendix E — Certified Host-Vector 
Systems 
While many experiments using E. coli 
K-12, Saccharomyces cerevisiae and 
Bacillus subti/is 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. sub til is HVl systems: 
pUBlia pCl94, pSl94. pSA2100, pEl9^, 
pTl27, pUBH2. pC221. pC223. and 
pABl24. B. subtilis strains RUB 331 and 
BGSC 1S53 have been certified as the 
host component of HVl systems based 
on these plasmids. 
HV2. The asporogenic mutant 
derivative of Bacillus subtilis. ASB 298. 
with the following plasmids as the 
vector component: pUBllO, pCl94, 
r 147] 
