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Federal Register / Vol. 51, No. 88 / Wednesday, May 7, 1986 / Notices 
Streptococcus milled 
Streptococcus durans 
Streptococcus mitior 
Streptococcus Ferus 
Exceptions. Experiments described in 
Section III— A which require specific 
RAC review and NIH approval before 
initiation of the experiment. 
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-VI — Footnotes end 
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 3iosafety, 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 3 to these 
Guidelines. 
2. A subset cf 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 under Section III— D— 4. 
4. As classified in the Third Report of the 
International Committee on Taxonomy of 
Viruses: Classification and Nomenclature of 
Viruses, R.E.F. Matthews. Ed. Interviro'ogy 12 
(129-296) 1979. 
5. i.e.. the total of ell genomes within a 
Family shall not exceed one-half of the 
genome. 
Appendix D — Actions Taken Under the 
Guidelines 
As noted in the subsections of Section 
IV- C-l-b-(l), the Director, NTH, 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 EK1 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-I I 
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 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 
Hemophilus plasmid, pRSF0885. under 
BLl conditions. 
Appendix D-IV 
Permission is granted to clone certain 
subgenomic segments of foot and mouth 
disease virus in HVl Bacillus subtilis 
and Sacchcromyces cerevisiae host- 
vector systems under ELI 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-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 
Sclmonella 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-VHI 
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 cf Cornell University to field 
test tomato and tobacco plants 
transformed with bacterial [E. ccli K-12) 
and yeast DNA using pollen as a vector. 
Appendix D-X 
Permission is granted to Drs. Steven 
Lindow and Nicholas Panopouios of the 
University of California, Berkeley, to 
release under specified conditions 
Pseudomoncs syringee pv. syringae and 
Erwinia herbicola carrying in vitro 
generated deletions of all or part of the 
genes involved in ice nucleation. 
Appendix D-Xl 
Agracetus of Middleton, Wisconsin, 
may field test under specified conditions 
disease resistant tobacco plants 
prepared by recombinant DNA 
techniques. 
Appendix E — Certified Host-Vector 
Systems 
(See also Appendix I) 
While many experiments using E. coli 
K-12, Saccharomyces cerevisiae and 
Bacillus subtilis are currently exempt 
from the Guidelines under Section III-D- 
5, some derivatives of these host-vector 
systems were previously classified as 
HVl. or HV2. A listing of those systems 
follows: 
Appendix E-I — Bccillus subtilis 
HVl. The following plasmids are 
accepted as the vector components of 
certified B. subtilis HVl systems: 
pUBllO, pCl94, pS!94, pSA2100, pEl94, 
pTl27, pUBll2, 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, pCl34, 
pSl94, pSA2100, pEl94, pT127. pUBl12, 
pC221, pC223, and pABl24. 
Appendix E-Il — Saccharomyces 
cerevisiae 
HV2. The following sterile strains of 
Saccharomyces cerevisiae, all of which 
have the ste-VC9 mutation, SHYl, 
SHY2, SHY3, and 3HY4. The following 
plasmids are certified for use: YIpl, 
YEp2, YEp4, YIp5, YEp0, YRp7, YEp20. 
YEp21. YEp24, YIp25, YIp26, YIp27, 
YIp28, YIp29, YIp30, YIp31, YIp32, and 
YIp33. 
Appendix E-III — Escherichia coli 
EX2 Plasmid Systems. The E. coli K- 
12 strain chi-1776. The followinn 
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