Federal Register / Vol. 51. No. 88 / Wednesday, May 7, 1986 / Notices 
16969 
Appendix C-I — Recombinant DMAs in 
Tissue Culture. 
Recombinant DNA molecules 
containing less than one-half of any 
eukaryotic genome (all viruses from a 
single Family (4) being considered 
identical (5)) that are propagated and 
maintained in cells in tissue culture are 
exempt from these Guidelines with the 
exceptions listed below. 
Exceptions. Experiments described in 
Section III— A which require specific 
RAC review and NIH approval before 
initiation of the experiment. 
Experiments involving DN’A from 
Class 3. 4. or S organisms [1] or cells 
known to be infected with these agents. 
Experiments involving the deliberate 
introduction of genes coding for (he 
biosynthesis of molecules toxic for 
vertebrates (see Appendix F). 
Appendix C-II — Experiments Involving 
E. coli K-12 Host - Vector Systems 
Experiments which use E. coli K-12 
host-vector systems, with the exception 
of those experiments listed below, are 
exempt from these Guidelines provided 
that: (i) the E coli host shall not contain 
conjugation proficient plasmids or 
generalized transducing phages: and (ii) 
lambda or lambdoid or Ff 
bacteriophages or nonconjugative 
plasmids (2] shall be used as vectors. 
However, experiments involving the 
insertion into E. coli K-12 of DNA from 
prokaryotes that exchange genetic 
information (3) with E. coli may be 
performed with any E coli K-12 vector 
(e g . conjugative plasmid). When a 
nonconjugative vector is used, the E. 
coli K-12 host may contain conjugation- 
proficient plasmids either autonomous 
or integrated, or generalized transducing 
phages. 
For these exempt laboratory 
experiments. BLl physical containment 
conditions are recommended. 
For large-scale (LS) fermentation 
experiments BLl-LS physical 
containment conditions are 
recommended. However, following 
review by the IBC of appropriate data 
for a particular host-vector system, some 
latitude 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 L1I-A which require specific 
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 III— B— 2 with prior IBC review 
and approval. 
Recombinant DNA Research, 
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-Ill — Experiments Involving 
Saccbaromyces Host-Vector Systems 
Experiments which use 
Saccbaromyces cerevisiae host-vector 
systems, with the exception of 
experiments listed below, are exempt 
from these Guidelines. 
Experiments which use 
Saccbaromyces uvarum host-vector 
systems, with the exception of 
experiments listed below, are exempt 
from these Guidelines. 
For these exempt laboratory 
experiments. BLl physical containment 
conditions are recommended. 
For large-scale fermentation 
experiments BLl-LS physical 
containment conditions are 
recommended. However, following 
review by the IBC of appropriate data 
for a particular host-vector system some 
latitude in the application of BLl-LS 
requirements as outlined in Appendix 
K-II-A through K— II— F is permitted. 
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 knowns 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-TV— Experiments Involving 
Bacillus subtilis Host- Vector Systems 
Any asporogenic Bacillus subtilis 
strain which does not revert to a 
sporefermer with a frequency greater 
than 10" 7 can be used for cloning DNA 
with the exception of those experiments 
listed below. 
For these exempt laboratory 
experiments. BLl physical containment 
conditions are recommended. 
For large-scale fermentation 
experiments BLl-LS physical 
containment conditions are 
recommended. However, following 
review by the IBC of appropriate data 
for a particular host-vector system, some 
Volume 1 1 
latitude in the application of BLl-LS 
requirements as outlined in Appendix 
K-II-A through K-II-F is permitted. 
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 — Extrccbromosomal 
Elements of Cram Positive Organisms 
Recombinant DN’A 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 Guidelines. 
Bacillus subtilis 
Bacillus pumilus 
Bacillus licheniformis 
Bacillus thuringiensis 
Bacillus cereus 
Bacillus amyloliquefaciens 
Bacillus brevis 
Bacillus natto 
Bacillus niger 
Bacillus atem'mus 
Bacillus amylosacchariticus 
Bacillus anthracis 
Bacillus globigii 
Bacillus megaterium 
Staphylococcus aureus 
Staphylococcus epidermidis 
Staphylococcus camosus 
Clostridium acetobutylicum 
Pediococcus damnosus 
Pediococcus pentosaceus 
Pediococcus acidilactici 
Lactobacillus casei 
Listeria grayi 
Listeria murrayi 
Listeria monocytogenes 
Streptococcus pyogenes 
Streptococcus agalactiae 
Streptococcus sanguis 
Streptococcus salivarious 
Streptococcus cremoris 
Streptococcus pneumoniae 
Streptococcus avium 
Streptococcus faecalis 
Streptococcus anginosus 
Streptococcus sobrinus 
Streptococcus lactis 
Streptococcus mutans 
Streptococcus equisimilis 
Streptococcus thermophylus 
[13] 
