7184 
Federal Register / Vol. 45, No. 22 / Thursday, January 31, 1980 / Notices 
the cloning of the exotoxin A gene of 
Pseudomonas aeruginosa in E. coli K-12 
is permitted. Under exemption I-E-4, P. 
aeruginosa and genus Escherichia are 
listed in Appendix A, Sublist A. 
However, according to Section I-E, 
Exemptions: 
It must be emphasized that the following 
exemptions (4) are not meant to apply to 
experiments described in the Sections I-D-l 
to I-D-5 as being prohibited. 
Section l-D-2 prohibits: 
Deliberate formation of recombinant DNAs 
containing genes for the biosynthesis of 
toxins potent for vertebrates (2A) (e.g., 
botulinum, diphtheria toxins; venoms from 
insects, snakes, etc). 
Dr. Shuster notes that exotoxin A and 
P. aeruginosa are safe under normal 
laboratory conditions. P. aeruginosa is 
an opportunistic pathogen and exotoxin 
A is only one of a series of virulence 
factor necessary to produce 
pathogenicity. This request was 
presented to the RAC at its December 6- 
7, 1979 meeting. At that time, the RAC 
felt that approval of the proposed 
experiments appeared to require an 
exception to a prohibition. Accordingly, 
the request is being published in the 
Federal Register for a 30 day comment 
period prior to being reconsidered by 
the RAC at its March 6-7, 1980 meeting. 
11. Request for Consideration of 
Appropriate Containment Levels. Dr. 
Marvin Schwalb of New Jersey Medical 
School has requested that the RAC 
consider the containment levels 
appropriate to the return of 
Schizophyllum commune DNA cloned in 
Saccharomyces cerevisiae to 
Schizophyllum commune. In addition, 
Dr. Schwalb requests permission to 
clone the S. cerevisiae derived vector 
YR414/ura 3 and the Saccharomyces 2 
plasmid containing yeast or S. commune 
sequences in S. commune. 
2. Request for Consideration of 
Appropriate Containment Levels. Dr. 
Olen Yoder of Cornell University has 
asked the RAC to consider the 
appropriate containment level for the 
return of Helminthosporanium maydis 
DNA, which has been cloned in 
Saccharomyces cerevisiae, to the host 
of origin. 
13. Request to Lower Containment 
Conditions for Prokaryotic Cloning. Dr. 
David Wilson of Cornell University has 
requested a lowering from P3 to P2 
containment levels for experiments 
involving the cloning of the cellulase 
gene of Sporocytophaga ep. into a 
thermophilic Bacillus. 
14. Recombinant DNA Experiments 
Involving Foot-And-Mouth Disease 
Virus. The RAC at its December 6-7, 
1979 meeting recommended, and the 
Director, NIH, subsequently approved, 
the cloning of foot-and-mouth disease 
virus in E. coli K-12 at the Plum Island 
Animal Disease Center. The RAC 
further recommended, and the Director, 
NIH, subsequently approved, that prior 
to the shipment of any clones off of Plum 
Island, first a working group of the RAC 
and subsequently the full RAC would 
examine data arising from the foot-and- 
mouth disease virus recombinant DNA 
work on Plum Island. 
If any such data are ready for 
presentation to the RAC, they will be 
considered at the March 6-7, 1980 
meeting. The RAC analysis may include 
reexamination of the appropriate 
containment levels for experiments with 
any clones recommended to be shipped 
from Plum Island. 
15. Proposed EK2 Host- Vector 
Systems. Dr. P. H. Pouwels of the 
Mediach Biologisch Laboratorium of 
Rijswijk, The Netherlands, has 
requested EK2 certification of several 
trp plasmids derived from plasmids 
pBR345 and pBR313. The tip vectors 
derived from pBR345 carry no antibiotic 
resistance genes. These plasmids would 
be used in conjunction with the host 
xl776 as an EK2 certified host-vector 
system. 
16. Revision of Section 111-C-l-e of 
the Guidelines. The proposed revised 
Guidelines (44 FR 69210), in Appendix C, 
cite a limited exemption, under Section 
I-E-5, for certain recombinant DNA 
experiments in tissue culture: 
Under exemption I-E-5 of these Revised 
Guidelines are 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.). 
This exemption was promulgated in 
the Federal Register of July 20, 1979 (44 
FR 42914). In its original formulation (44 
FR 22314), the exemption proposed to 
include recombinant DNA molecules 
that contain no more than one-fourth of 
the genome of a eukaryotic virus. At its 
December 6-7, 1979 meeting, Dr. 
Wallace Rowe submitted a document 
requesting that the RAC reconsider the 
question of allowing exemption for 
tissue culture experiments involving 
recombinant molecules containing no 
more than one-fourth of the genome of a 
virus. A working group was appointed to 
make recommendations for 
consideration at the March 6-7, 1980. 
meeting. It has been recommended that 
Sections III— C— 1— e, III— C— 1 c (1), III— C— 
1— e— (1)— (a), and III— C— 1— e— (1)— (b) of the 
Guidelines be changed and that a new 
Section UI-C-l-e-(l)-(c) be added. 
Section III-C-l-e-(2) would remain 
unchanged. The proposed revised 
sections would read as follows: ( 
m-C-l-e. All Viral Vectors. 
III-C-l-e-{l). Other experiments involving 
eukaryotic virus vectors can be done as 
follows: 
IU-C-l~e-(l)-(a). Recombinant DNA 
molecules containing no more than two-thirds 
of the genome of any eukaryotic virus (all 
viruses from a single Family being considered 
identical) may be propagated and maintained 
in cells in tissue culture in the absence of 
helper virus using Pi containment. The DNA 
may contain fragments of the genomes of 
viruses from more than one Family but each 
fragment must be less than two-thirds of a 
genome. For such experiments, no MUA need 
be submitted but prior notice must be given 
to the IBC as described in Section III— 0 of the 
Guidelines. The IBC should handle such 
registration documents as described in 
Section ID-0. 
‘ 'III-C-l-e-( l)-(b ). Recombinants with less 
than two-thirds of the genome of any 
eukaryotic virus may be rescued with helper 
virus using P2 containment if wild type 
strains of the helper virus are not able to 
grow in human cells. 
III-C-1 -e-{ l)-(c). Recombinants with less 
than two-thirds of the genome of any 
eukaryotic virus may be rescued with helper 
virus using P3 containment if wild-type 
strains of the helper virus are able to grow in 
human cells. 
17. Consideration of Appropriate 
Containment Levels. Dr. Charles Jacobs 
of the University of Texas at Austin has 
requested permission from the RAC to 
clone Wangiella dermatitidis DNA in 
Wangiella dermatitidis using 
Saccharomyuces/E. coli hybrid 
plasmids as vectors. Dr. Jacobs requests 
that the RAC assess appropriate 
containment conditions. 
Dated: January 23, 1980. 
Donald S. Fredrickson, M.D., 
Director, National Institutes of Health. 
[FR Doc. 80-3041 Filed 1-30-80: 8:45 am) 
BILLING CODE 4110-08-M 
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