1M56 
Federal Regletar / Vol. 46. No. 46 / Tbunday, March 12, 1981 / Notices 
•ubsectiona of Part m. including ID-O-1. 
ni-O-2. etc." 
ll-B. Addition of New Paragraph to Part 
III 
A new paragraph is added just before 
Section Hl-O beg^. as follows: 
“When the reader Qnds that the 
containment level given for the same 
experiment is different in two different 
sections within Part III. he may choose 
whichever of the two levels he wishes to 
use for the experiment" 
/f-C Addition of New Section DI-0~2 
A new section. m-O-2, is added to 
the guidelines as follows: 
"1II-0-2. Experiments Involving 
Prokaryotes Nonpathogenic for Man, 
Animals or Plants, ana/or Lower 
Eukaryotes Nonpathogenic for Man. 
Animals or Plants. Recombinant DNA 
experiments involving prokaryotes 
nonpathogenic for man. animals, or 
plants, and/or lower eukaryotes 
nonpathogenic for man. animals or 
plants, and only DNA from such 
sources, can be conducted under P3 
containment (2A). Lower levels of 
ysical containment may be assigned 
ORDA on a case-by-case basis for 
spedflc donor-recipient combine tloiu 
(see Section rV-E-l-b-(3Hh))." 
II-D. Amendment of Section lU-B-3 
The second paragraph of Section ID- 
B-3 is deleted. Revised Section ID-B-3, 
now reads as foUows; 
“ID-B-3. Non-HVJ Systems. 
Containment levels for other classes of 
experiments involving non-HVL systems 
may be approved by the Director, NIH. 
(See Sections fV-E-l-b-flHb). IV-E-1- 
b-(2Hc). IV-E-l-b-{3Hb)).- 
U-E Deletion of Section rV-E-l-b~(2)- 
(f) 
Section IV-E-l-b-(2Hn !• deleted. 
U-P. Addition of Section lV-E-l-b-(3)- 
(h) 
A new section IV-E-l-b-(3Hh) is 
added as follows: 
’TV-Er-l-b-(3Hh), Assigning 
containment Iwala for experiments in 
which both donor and recipient are 
nonpathogenic prokaryotes and/or 
nonpathogenic lower eukaryotes (see 
Section ni-O-2)." 
H-C. Amendment of Appendix A, 
Sublist F 
Appendix A. Sublist F is amended to 
read as follows: 
'Sublist P 
1 . Streptococcus sanguis 
2. Streptococcus pneumoniae 
3. Streptococcus foecalis” 
II-H. Amendments of Appendix E 
Entires 4 and 18 of Appendix E, are 
amended to read as follows: 
“4. Qoned desired fragments from any 
non-prohiblted source may be 
transferred Into Agrobacterium 
tumefaciens containing a Ti plasmid (or 
derivatives thereof), using a 
nonconjugatlve E coli plasmid vector 
coupled to a fragment of the TI plasmid 
and/or the origin of replication of an 
Agrobacterium plasmid, under 
containment conditions one step higher 
than would be required for the desired 
DNA in HVl systems (i.e.. one step 
higher than that specified in the 
subsections of Section lll-A). However, 
DNA from plants and nonpathogenic 
prokaryotes may be cloned uiuler P2 
containment conditions; and the 
Soccharomyces cerevisiae alcohol 
dehydrogenase 1 gene and the gene 
coding for the maize [Zea mays) seed 
storage protein, zein, may be cloned 
under Pi conditions. Transfer Into plant 
parts or cells in culture is permitted at 
the same containment level as is used 
for the cloning in Agrobacterium 
tumefaciens. 
“18. DNA from nonpathogenic 
prokaryotes and nonpathogenic lower 
eukaryotes may be cloned into 
Schizosaccharomyces pombe species 
under Pi contaimnent conditions. DNA 
from higher eukaryotes may be cloned in 
S pombe species under P3 containment 
conditions." 
The following new entries are added 
to Appendix E: 
“28. Soccharomyces cerevisiae DNA 
may be cloned in Tetrahymena 
thermophila using E coli/E cerevisiae 
hybrid plasmids under Pi containment 
conditions." 
“Z7. All members of the 
nonpathogenic Actinomycetes genus 
Streptomyces and the plasmids native to 
this genus are approved as host-vector 
systems for the cloning under Pi 
conditions of DNA derived from other 
nonpathogenic prokaryotic organisms 
such as Streptomyces and other 
nonpathogenic Actinomycetes spedes, 
Escherichia coli K-12, Bacillus subtilis. 
Bacillus lichenformis. Bacillus 
circulans, and other nonpathogenic 
Bacillus species, and for the doning of 
DNA derived from nonpathogenic 
unicellular eukaryotic ^croorganisms 
such as Soccharomyces cerevisiae and 
Neurosporo crassa . " 
//-/. Procedures for Review of Large- 
Scale Experiments 
The following procedures, which are 
not incorporate into the Guidelines, 
have been adopted for the review of 
large-scale exj^ments. (They are an 
expansion of the procediues which 
appeared in the November 21, 1980 
Federal Register (45 FR 77380)); 
“Application Procedures for Large- 
Scale Recombinant DNA Experiments. 
"1. For each research project 
proposing to exceed the 10-liter limit the 
applicant shall file a request with the 
NIH Office of Recombinant DNA 
Activities (ORDA). The request should 
include the following information; 
"a. The registration document 
submitted to the local Institutional 
Biosafety Committee (see Section III of 
the Guidelines). This should include, or 
have appended to it, a summary 
paragraph which describes the proposed 
project in language that is 
comprehensible to non-spedalists. 
"b. A statement of the rationale for 
wishing to exceed the 10-liter limit. 
"& Evidence that the recombinant 
DNAs to be employed in the research 
have been rigorously characterized and 
are free of harmful sequences. 
"d. Specification of the P-LS level 
proposed to be used as defined in the 
NIH Physical Contaimnent 
Recommendations for Large-Scale Uses 
of Organisms Containing Recombinant 
DNA Molecules (Federal Register, April 
11, 1880). 
“2. Each request submitted to ORDA 
shall be referred to a working group of 
the NIH Recombinant DNA Advisory 
Committee for review. 
“3. Following review and approval by 
the working group, each request shall 
submitted to the entire Recombinant 
DNA Advisory Committee for review. 
“4. Following review and approval by 
the RAC, each request shall be 
submitted to the Director, NIH. for the 
final review. 
“5. Applications for large-scale 
experiments which are submitted by 
institutions not receiving NIH funds for 
recombinant DNA research shall be kept 
confidential (provided the Institutions so 
desire) in accordance with the 
provisions of the NIH Guidelines for 
Research Involving Recombinant DNA 
Molecules and to the extent permitted 
by law. 
'*8. Proposals that the submitter 
considers to represent minor 
modifications of already approved 
experiments will be handled by an 
expedited procedure. A request must be 
submitted to ORDA. This request should 
include the changes made, the way in 
which these changes were made (e.g., 
mutagenesis, recloning), and the nature 
and results of any tests done to 
determine that no major change has 
occurred (e.g.. restriction enzyme 
analjrsis, tests for produced products, 
teets of vector mobilization). ORDA will 
( 69 ) 
