Federal Register / Vol. 59, No. 127 / Tuesday, July 5, 1994 / Notices 
34521 
pathogen or a plant pest; or (ii) the 
recombinant DNA codes for a toxin or 
other factor directly involved in 
eliciting human, animal, or plant 
disease or inhibiting plant growth, and 
is carried on an expression vector or 
wthin the host chromosome and/or 
when the host organism contains a 
conjugation proficient plasmid or a 
generalized transducing phage; or (iii) 
the recombinant DNA comes from a host 
organism or virus regulated as a human 
or animal pathogen or as a plant pest 
and has not been adequately 
characterized to demonstrate that it does 
not code for a factor involved in 
eliciting human, animal, or plant 
disease. 
Appendix H-III. Footnotes and 
References of Appendix H 
For further information on shipping 
etiologic agents contact: (i) The Centers 
for Disease Control and Prevention, 
ATTN; Biohazards Control Office. 1600 
Clifton Road, Atlanta, Georgia 30333, 
(404) 639-3883, FTS 236-3883; (ii) The 
U.S. Department of Transportation, 
ATTN: Office of Hazardous Materials 
Transportation. 400 7th Street, S.W., 
Washington, DC 20590, (202) 366-4545; 
or (iii) U.S. Department of Agriculture. 
ATTN; Animal and Plant Health 
Inspection Service, Import-Export 
Products, Room 756, Federal Building, 
6505 Belcrest Road, Hyattsville, 
Maryland 20782; for Animal Pathogens 
call (301) 436-7885; for Plant Pests 
(301) 436-6799. 
Appendix I. Biological Containment 
(See Appendix E) 
Appendix I-i. Levels of Biological 
Containment 
In consideration of biological 
containment, the vector (plasmid, 
organelle, or virus) for the recombinant 
DNA and the host (bacterial, plant, or 
animal cell) in which the vector is 
propagated in the laboratory will be 
considered together. Any combination 
of vector and host which is to provide 
biological conUtixunent shall be chosen 
or constructed so that the following 
types of "escape” ere minimized: (i) 
survival of the vecior in its host outside 
the laboratory, and (ii) transmission of 
the vecior from the propagation host to 
other non-laboratory hosts. The 
following levels of biological 
containment (host-vector systems) for 
prokaryotes are established. Appendices 
1-I-A through 1-U-B describe Levels of 
biological containment (host-vector 
systems) for prokaryotes. Specific 
criteria will depend on tlie organisms to 
be used. 
Appendix I-I-A. Host-Vector 1 Systems 
Host-Vector 1 systems provide a 
moderate level of containment. Specific 
Host-Vector 1 systems are: 
Appendix I-I- A-1 . Escherichia coli K- 
12 Host-Vector 1 Systems (EKl). The 
host is always Escherichia coli K-1 2 or 
a derivative thereof, and the vectors 
include non-conjugative plasmids (e g., 
pSClOl, ColEl, or derivatives thereof 
(see Appendices I-IH-A through G) and 
variants of bacteriophage, such as 
lambda (see Appendices I-ITl-H through 
0). The Escherichia coli K-12 hosts 
shall not contain conjugation-proficient 
plasmids, whether autonomous or 
integrated, or generalized transducing 
phages. 
Appendix I-I-A-2. Other Host-Vector 
1 Systems. At a minimum, hosts and 
vectors shall be comparable in 
containment to Escherichia coli K-12 
with a non-conjugative plasmid or 
bacteriophage vector. Appendix I-II 
describes the data to be considered and 
mechanism for approval of Host-Vector 
1 systems. 
Appendix I-I-B. Host-Vector 2 Systems 
Host-Vector 2 Systems provide a high 
level of biological contaixunent as 
demonstrated by data from suitable tests 
performed in the laboratory. Escape of 
the recombinant DNA either via survival 
of the organisms or via transmission of 
recombinant DNA to other organisms 
should be <1/10* under specified 
conditions. Specific Host-Vector 2 
systems are: 
Appendix I-I-B-1. For Escherichia 
coli K-12 Host-Vector 2 systems (EK2) 
in which the vector is a plasmid, no 
more than 1/10* host cells shall 
perpetuate a cloned DNA fragment 
under the specified non-permissive 
laboratory conditions designed to 
represent the natural environment, 
either by survival of the original host or 
as a consequence of transmission of the 
cloned DNA fragment 
Appendix I-l-B-2. For Escherichia 
coli K-12 Host-Vector 2 systems (EK2) 
in which the vector is a phage, no more 
than 1/10® phage particles shall 
perpetuate a clon^ DNA fragment 
under the specified non-permissive 
laboratory conditions designed to 
represent the natural environment, 
either as a prophage (in the inserted or 
plasmid form) In the laboratory host 
used for phage propagation, or survival 
in natural enviroruncnts and 
transferring a cloned DNA fragment to 
other hosts (or their resident prophages). 
Appendix I-II. Certification of Host- 
Vector Systems 
Appendix l-U-A. Responsibility. Host- 
Vector 1 systems (other than Escherichia 
coli K-12) and Host-Vector 2 systems 
may not be designated as such until 
they have been certified by the NIH 
Director. Requests for certification of 
host-vector systems may be submitted to 
the Office of Recombinant DNA 
Activities, National Institutes of Health, 
Building 31, room 4B11, Bethesda, 
Maryland 20892, (301) 496-9838. 
Proposed host-vector systems will be 
reviewed by the RAC (see section IV-C- 
l-b-(l)-(e)). Initial review will based on 
the construction, properties, and testing 
of the proposed host-vector system by a 
subcommittee composed of one or more 
RAC members and/or ad hoc experts. 
The RAC will evaluate the 
subcommittee's report and any other 
available information at the next 
scheduled RAC meeting. The NIH 
Director is responsible for certification 
of host-vector systems, following advice 
of the RAC. Minor modifications to 
existing host-vector systems (i.e., those 
that are of minimal or no consequence 
to the properties relevant to 
containment), may be certified by the 
NIH Director without prior RAC review 
(see section IV-C-l-b-(2)-(h)). Once a 
host-vector system has been certified by 
the NIH Director, a notice of 
certification will be sent by NIH/ORDA 
to the applicant and to the Institutional 
Biosafety Cominittee Chairs. A list of all 
currently certified host -vector systems is 
available from the Office of 
Recombinant DNA Activities, National 
Institutes of Health, Biylding 31, room 
4B11, Bethesda, Maryland 20892, (301) 
496-9838. The NIH Director may 
rescind the certification of a host-vector 
system (see section fV-C-l-b-(2)-(i)). If 
certification is rescinded, NIH will 
instruct investigators to transfer cloned 
DNA into a different system or use the 
clones at a higher level of physical 
containment level, unless NIH 
determines that the already constructed 
clones incorporate adequate biological 
containment Certification of an host- 
vector system does not extend to 
modifications of either the host or 
vector component of that system. Such 
modified systems shall be 
independently certified by the NIH 
Director. If modifications are minor, it 
may only be necessary for the 
investigator to submit data showing that 
the modifications have either improved 
or not impaired the major phcnot}q>ic 
traits on which the containment of the 
system depends. Substantial 
modifications to a certified host-vector 
system requires submission of complete 
testing data. 
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