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Federal Register / Vol. 59, No. 127 / Tuesday, July 5, 1994 / Notices 
Appendix C-III. Saccharomyces Host- 
Vector Systems 
Experiments involving 
Saccharomyces cerevisiae and 
Saccharomyces uvarum host-vector 
systems, with the exception of 
experiments listed in Appendix C-IIl-A, 
are exempt from the NIH Guidelines. 
For these exempt experiments, BLl 
physical containment is recommended. 
For IcLTge scale fermentation 
experiments, the appropriate physical 
containment conditions need be no 
greater than those for the host organism 
unmodified by recombinant DNA 
techniques; the Institutional Biosafety 
Committee can specify higher 
containment if deemed necessary. 
Appendix C-III-A. Exceptions 
The following categories are not 
exempt from the NIH Guidelines: (i) 
Experiments described in Section IIl-A 
which require Institutional Biosafety 
Committee approval, RAC review, and 
NIH approval before initiation, (ii) 
experiments described in Section Ill-B 
which require Institutional Biosafety 
Committee and NIH/ORDA approval 
before initiation, (iii) experiments 
involving DNA from Class 3, 4, or 5 
organisms (see Appendix C-Vl-A) or 
cells known to be infected with these 
agents may be conducted under 
containment conditions specified in 
Section IlI-C-2 with prior Institutional 
Biosafety Committee review and 
approval, (iv) large scale experiments 
(e.g., more than 10 liters of culture), and 
(v) experiments involving the deliberate 
cloning of genes coding for the 
biosynthesis of molecules toxic for 
vertebrates (see Appendix F). 
Appendix C-FV. Bacillus subtilis or 
Bacillus licheniformis Host-Vector 
Systems 
Any asporogenic Bacillus subtilis or 
asporogenic Bacillus licheniformis 
strain which does not revert to a spore- 
former with a frequency greater than 
10"’ may be used for cloning DNA with 
the exception of those experiments 
listed in Appendix C-FV-A. For these 
exempt laboratory experiments, BLl 
physical containment conditions are 
recommended. For large scale 
fermentation experiments, the 
appropriate physical containment 
conditions need be no greater than those 
for the host organism unmodified by 
recombinant DNA techniques: the 
Institutional Biosafety Committee can 
specify higher containment if it deems 
necessary. 
Appendix C-IV-A. Exceptions 
The following categories are not 
exempt from the NIH Guidelines; (i) 
Experiments described in Section III-A 
which require Institutional Biosafety 
Committee approval, RAC review, and 
NIH approval before initiation, (ii) 
experiments described in Section III-B 
which require Institutional Biosafety 
Committee and NIH/ORDA approval 
before initiation, (iii) experiments 
involving DNA from Class 3, 4. or 5 
organisms (see Appendix C-VI-A) or 
cells known to be infected with these 
agents may be conducted under 
containment conditions specified in 
Section III-C-2 with prior Institutional 
Biosafety Committee review and 
approval, (iv) large scale experiments 
(e.g., more than 10 liters of culture), and 
(v) experiments involving the deliberate 
cloning of genes coding for the 
biosynthesis of molecules toxic for 
vertebrates (see Appendix F). 
Appendix C-V. Extrachromosomal 
Elements of Gram Positive Organisms 
Recombinant DNA molecules derived 
entirely from extrachromosomal 
elements of the organisms listed below 
(including shuttle vectors constructed 
from vectors described in Appendix C), 
propagated and maintained in 
organisms listed below are exempt from 
these NIH Guidelines. 
Bacillus amyloliquefaciens 
Bacillus amylosacchariticus 
Bacillus anthracis 
Bacillus aterrimus 
Bacillus brevis 
Bacillus cereus 
Bacillus globigii 
Bacillus licheniformis 
Bacillus megaterium 
Bacillus natto 
Bacillus niger 
Bacillus pumilus 
Bacillus sphaericus 
Bacillus stearothermophilis 
Bacillus subtilis 
Bacillus thuringiensis 
Clostridium acetobutyliciim 
Lactobacillus casei 
Listeria grayi 
Listeria monocytogenes 
Listeria murrayi 
Pediococcus acidilactici 
Pediococcus damnosus 
Pediococcus pentosaceus 
Staphylococcus aureus 
Staphylcoccus carnosus 
Staphylococcus epidermidis 
Streptococcus agalactiae 
Streptococcus anginosus 
Streptococcus avium 
Streptococcus cremoris 
Streptococcus dorans 
Streptococcus equisimilis 
Streptococcus faecalis 
Streptococcus ferus 
Streptococcus lactis 
Streptococcus ferns 
Streptococcus mitior 
Streptococcus mutans 
Streptococcus pneumoniae 
Streptococcus pyogenes 
Streptococcus salivarious 
Streptococcus sanguis 
Streptococcus sobrinus 
Streptococcus thermophylus 
Appendix C-V-A. Exceptions 
The following categories of 
experiments are not exempt from the 
NIH Guidelines: (i) Experiments 
described in Section III-A which 
require Institutional Biosafety 
Committee, specific RAC review, and 
NIH approval before initiation, (ii) 
experiments described in Section III-B 
which require Institutional Biosafety 
Committee and NIH/ORDA approval 
before initiation, (iii) experiments 
involving DNA from Class 3, 4, or 5 
organisms (see Appendix C-VI-A) or 
cells known to be infected with these 
agents may be conducted under 
containment conditions specified in 
Section IIl-C-2 with prior Institutional 
Biosafety Committee review and 
approval, (iv) large scale experiments 
(e.g., more than 10 liters of culture), and 
(v) experiments involving the deliberate 
cloning of genes coding for the 
biosynthesis of molecules toxic for 
vertebrates (see Appendix F). 
Appendix C-Vl. Footnotes and 
References of Appendix C 
Appendix C-VI-A. 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 and Prevention, Office of 
Biosafety, Atlanta, Georgia 30333. 
Appendix C-VI-A-1. The NIH 
Director, with advice of the RAC. may 
revise the classification for the purposes 
of these NIH Guidelines (see Section IV- 
C-l-tM2)-(d)). The revised list of 
organisms in each class is reprinted in 
Appendix B. 
Appendix C-Vl-B. A subset of non- 
conjugative plasmid vectors are poorly 
mobilizable (e.g., pBR322, pBR313). 
Where practical, these vectors should be 
employed. 
Appendix C-VI-C. 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-E-5. 
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