Federal Register / Vol. 44, No. 141 / Friday, July 20, 1979 / Notices 
42917 
b. Approve introducing the bacteria 
into plant parts or cells in culture under 
P3 containment conditions. 
The motion was passed by the RAC 
by a vote of 14 for. 2 opposed, with 3 
abstentions. It was noted that this 
recommendation is narrower and more 
restrictive than the proposal published 
for comment in the Federal Register of 
April 13, 1979. It was also noted that 
recommendation of this proposal should 
not be construed as a general approval 
of the Agrobacterium system as a new 
cloning system. 
II. Summary of Major Actions Under 
Guidelines 
A. Containment levels for certified 
Saccharomyces cerevisiae and 
Neurospora crassa HV systems 
In accord with Section III— C— 5, host- 
vector systems which have been 
approved as HVl systems may be used 
under P2 containment conditions for 
shotgun experiments with phages, 
plasmids, and DNA from nonpathogenic 
prokaryotes which do not produce 
polypeptide toxins (34). For other 
classes of recombinant ONA 
experiments with these HVl systems, 
except for the cloning of complete 
genomes of eukaryote viruses the S. 
cerevisiae and N. crassa HVl systems 
and S. cerevisiae HV2 systems may be 
used at the physical containment levels 
applicable to EKl and EK2 systems, 
respectively. Experiments involving 
complete genomes of eukaryote viruses 
will require P3 + HV1 or P2 + HV2 
containment levels. 
B. Containment levels for unmodified 
laboratory strains of Neurospora crassa 
Unmodified laboratory strains of 
Neurospora crassa can be used in all 
experiments for which HVl N. crassa 
systems are approved provided that 
these are carried out at physical 
containment one level higher than 
required for HVl. However, if P3 
containment is specified for HVl N. 
crassa, this level is considered adequate 
for unmodified N. crassa. For P2 
physical containment, special care must 
be exercised to prevent aerial dispersal 
of macroconidia, including the use of a 
biological safety cabinet. 
C. III-C-7. Transfer of cloned DNA 
segments to Eukaryotic organisms 
III-C-7-a. Transfer to Non-human 
Vertebrates. DNA from any 
nonprohibited source [Section I-D], 
except for greater than one quarter of a 
eukaryotic viral genome, which has 
been cloned and propagated in E. coli 
under appropriate physical containment 
conditions, may be transferred with the 
E. coli vector used for cloning to any 
eukaryotic cells in culture or to any non- 
human vertebrate organism and 
propagated under conditions of physical 
containment comparable to Pi and 
appropriate to the organism under study 
[2AJ. Transfers to any other host will be 
considered by the RAC on a case-by- 
case basis [45]. 
Ill— C— 7— b. Transfer to Higher Plants. 
DNA from any nonprohibited source 
[Section I-D) which has been cloned 
and propagated in E. coli under 
appropriate containment conditions, 
may be transferred with the E. coli 
vector used for cloning to any higher 
plant organisms (Angiosperms and 
Gymnosperms) and propagated under 
conditions of physical containment 
comparable to Pi and appropriate to the 
organism under study [2Aj. Intact plants 
or propagative plant parts may be grown 
under Pi conditions described under 
Section III— C— 3. Containment must be 
modified to ensure that the spread of 
pollen, seed or other propagules is 
prevented. This can be accomplished by 
conversion to negative pressure in the 
growth cabinet or greenhouse or by 
physical entrapment by "bagging" of 
reproductive structures. Transfers to any 
other plant organisms will be considered 
on a case-by-case basis [45]. 
D. Exemption under I-E-5 for cloning in 
tissue culture cells 
In accord with Section I-E-5. those 
recombinant DNA molecules that are 
propagated and maintained in cells in 
tissue culture and that are derived 
entirely from non-viral components (that 
is. no component is derived from a 
eukaryotic virus) are exempt from the 
Guidelines. 
E. Containment levels for experiments 
involving genera Streptomyces and 
Micromonospora 
P2 physical containment shall be used 
for DNA recombinants produced 
between members of the genera 
Streptomyces and Micromonospora 
except for those species which are 
known to be pathogenic for man, 
animals or plants (2A). 
F. Exemption for Streptomyces species that 
exchange genetic information 
The following two sublists of 
Streptomyces species that have been 
shown to exchange chromosomal DNA 
are included under the exemption 
category of Section I-E-4 of the 1978 
Guidelines. Any recombinant DNA 
molecules that are composed entirely of 
DNA segments from one or more of the 
organisms within each sublist and to be 
propagated in any of the organisms 
included in that sublist are exempt from 
the Guidelines. (This list is to be 
separate from the other lists of exempt 
organisms in Appendix A.) 
Sublist 1 
Streptomyces aureofaciens 
Streptomyces rimosus 
Streptomyces coelicolor 
Sublis> 9 
Streptomyces griseus 
Streptomyces cyaneus 
Streptomyces venezuelae 
G. Use of Agrobacterium Tumefaciens 
as a host-vector system 
The NIH has approved the cloning of 
well-characterized fragments of 
eukaryotic DNA under P3 conditions, 
either in E. coli K-12 or in A. 
tumefaciens carrying a Ti plasmid, using 
an EK2 plasmid vector coupled to a 
fragment of the Ti plasmid and/or the 
origin of replication of a cryptic A. 
tumefaciens plasmid. 
The NIH has approved introducing 
these bacteria into plant parts or cells in 
culture under P3 containment 
conditions. 
Dated: July 13, 1979. 
Donald S. Fredrickson. 
Director. National Institutes of Health. 
|FR Doc. 79-12U0 Filed 7-1S-79; a«5 am| 
BILLING COOC 4110-M-M 
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