Federal Register / Vol. 49, No. 227 / Friday, November 23, 1984 / Notices 
46277 
Appendix C-IJI. Experiments 
Involving Sciccharomyces 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 laboratory 
experiments, BLl physical containment 
conditions are recommended. 
For large-scale fermentation 
experiments BLl-LS phystcial 
containment conditions are 
recommended. However, following 
review by the IBC or appropriate data 
for a particular host-vector system some 
latitute in the application of BLl-LS 
requirements as outlined in Appendix 
K-II-A through K-1I-F is permitted 
Exceptions. 
Experiments described in Section III— 
A which reqire specific RAC review and 
NIH approval before initiation of the 
experiment. 
Experiments involving Glass 3, 4 . or 5 
organisms(l) or cells known to be 
infected with these agents may be 
conducted under containment 
conditions specified in Section LU-B-2 
with prior IBC review and approval. 
Large-scale experiments (e-g.. more 
than 10 liters of culture) require prior 
IBC review and approved (see Section 
III-B-5). 
Experiments involving the deliberate 
cloning of genes coding for the 
biosynthesis of molecules toxic for 
vertebrates (see Appendix P). 
Appendix C-IV. Experiments 
Involving Bacillus subtilis Host-Vector 
Systems. Any a9porogenic Bacillus 
subtilis strain which does not revert to a 
sporeformer with a frequency greater 
than 10" 7 can be used foe cloning DNA 
with the exception of these 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 laboratory 
experiments, BLl physical containment 
conditions are recommended. 
For large-scale fermentation 
experiments BLl-LS physicial 
containment conditions are 
recommended. However, following 
reivew by the IBC of appropriate data 
for a particular host-vector system, 
sopme latitude in the application of BLl- 
LS requirements as oulined in Appendix 
K-II-A through K— II— F is permitted. 
Exceptions. 
Experiments described in Section HI— 
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 G-V — Footnotes and Reference* of 
Appendix C 
(/) 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, Georgia 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-con|ngative plasmid 
vectors are also poorly mobilizabte (e.g., 
pBR322. pBR313). Where practical, these 
vectors should be employed. 
(3) Defined as observable under optimal 
laboratory comditions 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 1II-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—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 9an Francisco, 
California. Further development of the 
clones at Genentech has been approved 
under BLl -+• EKl conditions. 
Appendix D-III. 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 
BLl conditions. 
Appendix D-FV. Permission is granted 
to clone certain subgenamic segments of 
foot and mouth disease virus in HVl 
Bacillus subtilis and Saccharomyces 
cerevisiae host-vector systems under 
BLl conditions at Genentech, 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 
techniques under specified containment 
conditions. 
Appendix D-VI. Permission is granted 
to clone in E. coli K-12 under BLl 
physical containment conditions 
subgenomic segments of rift valley fever 
virus subject to conditions which have 
been set forth by the RAC 
Appendix D-V II. Attenuated 
laboratory strains of Salmonella 
typhimurium may be used under BLl 
physical containment conditions to 
screen for the Saccharomyces 
cerevisiae pseudouridine synthetase 
gene. The plasmid YEpl3 will be 
employed as the vector. 
Appendix D-Vlll. 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 BLl 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 
PanopouJos 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 
(See also Appendix I.) 
While many experiments using E. colt 
K-12, Saccharomyces cerevisiae and 
Bacillus subtilis are currently exempt 
from the Guidelines under Section UI-D- 
5, some derivatives of these host-vector 
systems were previously classified as 
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