Federal Register / Vol. 46. No. Ill / Wednesday, June 10, 1981 / Notices 
30777 
appropriate animal model systems. 
Results will be published in the open 
scientific literature and will be 
presented in a subsequent update of the 
Final Plan. 
f. NIH scientists are also determining 
the biological activity of E. coli K-12 
clones carrying DNA copies of an RNA 
tumor virus. These studies involve the 
administration to hamsters of bacterial 
preparations containing Harvey 
sarcoma virus DNA ligated to lambda 
and pBR322. These studies are still in 
progress and when definitive results 
have been obtained they will be 
published in the open scientific 
literature and will appear in a 
subseauent update of the Final Plan. 
g. Although they had not appeared in 
the previous Final Plan, two additional 
NLAID contract-supported projects are 
peripherally related to the total risk 
assessement activities. 
(1) RAC had suggested that a model 
cloned DNA segment be constructed to 
serve as a uniform heterologous DNA 
segment for risk assessment studies. 
This endeavor has been successfully 
completed by an investigator at the 
University of Wisconsin and employed, 
as a source of DNA. the plasmid NR79. 
The EcoRl fragment NR79-R1-G1 
contains genes encoding for 
chloramphenicol, kanamycin and 
sulfonamide resistance, as well as the 
promoters that govern these resistances. 
NR79-R1-C1 was shown not to have an 
internal EcoRl site and also not to be 
capable of replicating by itself or with 
the help of other plasmids. Experimental 
results indicated that NR79-R1-G1 is not 
capable of autonomous replication and 
does not contain an origin of replication 
that can function in a recombinant 
plasmid. The antibiotic resistances 
encoded for by NR79-R1-G1 were tested 
and found not to be transposable, that is 
gene movement and insertion at a new 
site be a mechanism which does not 
involve the homologous recombination 
system of the host strain. 
Since the fragment is not capable of 
autonomous replication, plasmid pfT353. 
a recombinant plasmid containing 
NR79-R1-C1 cloned into the EcoRl site 
of pBR322. is the suggested way of 
propagating this fragment. Stocks of E. 
coU strain KH802 containing pJT353 
have been preserved for the propagation 
and isolation of NR79-R1-G1. Protocols 
used for the isolation of puriFied plasmid 
p|T353 (NR79-R1-C1 and pBR322j and 
puriFied NR79-R1-C1 fragment are 
available. 
This work was supported by Contract 
J'JPl-AI-QZeoe. University of Wisconsin. 
Robert Rownd. principal investigator. 
(2) A contract has been awarded to 
I the University of Minnesota. (Contract 
NOl-AI-02654. Donald Vesley, principal 
investigator) to develop a 
Comprehensive Course on 
Microbiological Principles and 
Techniques for Work with Potentially 
Biohazardous Agents Including 
Recombinant DNA. The Principal 
Investigator will work with the Board of 
Education and Training of the American 
Society for Microbiology (ASM) in 
developing the materials. 
In 1977, the ASM undertook a study to 
devise standards of training for 
recombinant DNA research workers. A 
proper state of training of the laboratory 
workers is the First line of containment 
and it was their opinion that such 
training standards should be set by 
knowledgeable professionals. The 
product of that study was not standards 
but rather an outline of a body of 
knowledge which it was felt that any 
Principal Investigator should be aware 
of before independently embarking on 
recombinant DNA research. This 
contract will develop all necessary 
resources to present a course based on 
the ASM Findings. 
The plan is for the NLAID to support 
the development of the resource 
materials and once that phase is 
completed, the NIH Division of Safety 
will have the materials reproduced and 
distributed to the various Institutions 
where the NIH supports research to 
assist them in performing their own 
local training responsibilities. We will 
devise both a standard lecture/ 
laboratory course and self study aides. 
2. Eukaryotic Host- Vector Systems. 
Lower eukaryotic and higher 
eukaryotic host/vector systems were 
given a lower priority in the Final Plan 
than prokaryotic systems. 
‘After discussion at both the March 
and June 1980 meetings, the RAC 
recommended modifying the Guidelines 
to include Saccharomyces cerevisioe 
host-vector systems under Section Ill-O. 
During their consideration the RAC 
considered information that; (1) S. 
cerevisioe is nonpathogenic. (2) it does 
not implant in the intestine. (3) the dilute 
conditions in which it is found in nature 
are extremely unfavorable for mating, 
(4) it does not efFiciently compete with 
wild strains of 5. cerevisioe, and (5) it is 
fully sensitive to autoclaving and 
disinfection by standard agents. 
RAC also considered the question of 
proper physical containment for higher 
eukaryotic viral vectors. The Guidelines 
have been modiFied to permit work at 
more relaxed levels of physical 
containment than previously required. 
The Guidelines have been modiFied to 
permit 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) to be propagated 
and maintained in cells in tissue culture 
using Pi containment. It must be shown 
that the cells lack helper virus for the 
speciFic families of the defective viruses 
being used. 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. 
No initiatives have been started to 
directly answer the issues raised in the 
Final Plan relative to eukaryotic host- 
vector systems. NIH will continue to 
monitor the results of free ranging 
research to collect useful data that can 
be analyzed as part of the risk 
assessment process. 
C. Implementation. 
The original Final Plan stated that 
NIAID would recruit and appoint an 
eminent scientist as a Special Assistant 
to the Director, NIAID for Risk 
Assessment. Attempts to recruit a 
person to Fill this position were made 
including national advertising. No 
individual with the desired level of 
credentials and broad recognition by the 
various scientiFic disciplines was 
identiFied who was willing to under take 
this work at the NIH under the 
conditions we could offer. By midyear it 
had become evident that the probable 
scope and number of various activities 
encompassed by the Risk Assessment 
Plan would probably stabilize at the 
current level and not increase further. 
Consequently, the necessity for a 
Special Assistant was reconsidered and 
alternatives for satisfying the needs 
were evaluated. A decision was made to 
distribute the proposed functions of the 
Special Assistant to the Office of 
Recombinant DNA Activities and the 
Office of Specialized Research and 
Facilities, both within the NIAID. When 
appropriate, these ofFices will use ad 
hoc consultants in fulfilling the tasks 
originally described for the Special 
Assistant and for which staff and the 
RAC Risk Assessment Subcommittee 
feel that additional or specific expertise 
is required. 
The remainder of the Implementation 
section of the plan remains in effect as 
staled. Most implementation actions 
have been cited earlier in this document. 
However, an important part of the Plan 
is the submission of periodic reports to 
the RAC. In this regard, the Director, 
NIAID, has reported to the RAC on 
activities related to risk assessment, and 
has provided thereby an important and 
continuing review of progress, and 
receives the advice or comment of this 
advisory group. 
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