Federal Register / Vol. 44, No. 232 / Friday, November 30, 1979 / Notices 
69217 
cloned DNA fragment under the 
specified nonpermissive 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. 
II-D-l-b-(2). For EK2 host-vector 
systems in which the vector is a phage, 
no more than one in 10* phage particles 
should be able to perpetuate a cloned 
DNA fragment under the specified 
nonpermissive laboratory conditions 
designed to represent the natural 
environment either (i) as a prophage (in 
the inserted or plasmid form) in the 
laboratory host used for phage 
propagation or (ii) by surviving in 
natural environments and transferring a 
cloned DNA fragment to other hosts (or 
their resident prophages). 
Il-D-l-c. HV3. These are host-vector 
systems in which: 
II— D— 1— c— (1). All HV2 criteria are met. 
Il-D-l-c-{2). The vector is dependent 
on its propagation host or is highly 
defective in mobilizability. Reversion to 
host-independence must be less than 1/ 
10* per vector genome per generation. 
II— D— 1— c— (3). No markers conferring 
resistance to antibiotics commonly used 
clinically or in agriculture are carried by 
I the vector, unless expression of such 
! markers is dependent on the 
propagating host or on unique 
laboratory-controlled conditions or is 
! blocked by the inserted DNA. 
II— D— 1— c— (4). The specified 
I containment shown by laboratory tests 
has been independently confirmed by 
i specified tests in animals, including 
| primates, and in other relevant 
environments. 
II-D-l-c-(5). The relevant genotypic 
and phenotypic traits have been 
independently confirmed. 
Il-D-2. Certification of Host- Vector 
' Systems. 
II-D-2-a. Responsibility. HVl 
systems other than E. coli K-12. and 
HV2 and HV3 host-vector systems, may 
* not be designated as such until they 
have been certified by the Director, NIH. 
Application for certification of a host- 
vector system is made by written 
application to the Office of Recombinant 
DNA Activities (ORDA). National 
Institutes of Health, Bethesda, Maryland 
20205. 
Host-vector systems that are proposed 
for certification will be reviewed by the 
NIH Recombinant DNA Advisory 
Committee (RAC). (See Section IV-E-1- 
t>— (1)— (c).) This will first involve review 
of the data on construction, properties, 
and testing of the proposed host-vector 
system by a Working Group composed 
of one or more members of the RAC and 
other persons chosen because of their 
expertise in evaluating such data. The 
Committee will then evaluate the report 
of the Working Group and any other 
available information at a regular 
meeting. The Director, NIH is 
responsible for certification after 
receiving the advice of the RAC. Minor 
modifications of existing certified host- 
vector systems, where the modifications 
are of minimal or no consequence to the 
properties relevant to containment may 
be certified by the Director, NIH without 
review by the RAC. (See Section IV-E- 
l-b-(3)-(f).) 
When new host-vector systems are 
certified, notice of the certification will 
be sent by ORDA to the applicant and to 
all IBCs and will be published in the 
Recombinant DNA Technical Bulletin. 
Copies of a list of all currently certified 
host-vector systems may be obtained 
from ORDA at any time. 
The Director, NIH may at any time 
rescind the certification of any host- 
vector system. (See Section IV-E-l-b- 
(3H0 ) If certification of a host-vector 
system is rescinded, NIH will instruct 
investigators to transfer cloned DNA 
into a different system, or use the clones 
at a higher physical containment level 
unless NIH determines that the already 
constructed clones incorporate adequate 
biological containment. 
Certification of a given system does 
not extend to modifications of either the 
host or vector component of that system. 
Such modified systems must be 
independently certified by the Director. 
NIH. 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 phenotypic traits 
on which the containment of the system 
depends. Substantial modifications of a 
certified system require the submission 
of complete testing data. 
II-D-2-b. Data To Be Submitted for 
Certification. 
II— D— 2— b— { 1 ). HVl Systems Other than 
E. Coli K-12. The following types of data 
shall be submitted, modified as 
appropriate for the particular system 
under consideration: (i) A description of 
the organism and vector; the strain's 
natural habitat and growth 
requirements; its physiological 
properties, particularly those related to 
its reproduction and survival and the 
mechanisms by which it exchanges 
genetic information; the range of 
organisms with which this organism 
normally exchanges genetic information 
and whet sort of information is 
exchanged; and any relevant 
information on its pathogenicity or 
toxicity, (ii) A description of the history 
of the particular strains and vectors to 
be used, including data on any 
mutations which render this organism 
less able to survive or transmit genetic 
information, (iii) A general description 
of the range of experiments 
contemplated, with emphasis on the 
need for developing such an HVl 
system. 
II— D— 2— b— (2). HV2 Systems. 
Investigators planning to request HV2 
certification for host-vector systems can 
obtain instructions from ORDA 
concerning data to be submitted [33A, 
33B]. In general, the following types of 
data are required: (i) Description of 
construction steps, with indication of 
source, properties, and manner of 
introduction of genetic traits, (ii) 
Quantitative data on the stability of 
genetic traits that contribute to the 
containment of the system, (iii) Data on 
the survival of the host-vector system 
under nonpermissive laboratory 
conditions designed to represent the 
relevant natural environment, (iv) Data 
on transmissibility of the vector and/or 
a cloned DNA fragment under both 
permissive and nonpermissive 
conditions, (v) Data on all other 
properties of the system which affect 
containment and utility, including 
information on yields of phage or 
plasmid molecules, ease of DNA 
isolation, and ease of transfection or 
transformation, (vi) In some cases, the 
investigator may be asked to submit 
data on survival and vector 
transmissibility from experiments in 
which the host-vector is fed to 
laboratory animals (e.g„ rodents). Such 
in vivo data may be required to confirm 
the validity of predicting in vivo survival 
on the basis of in vitro experiments. 
Data must be submitted in writing to 
ORDA. Ten to twelve weeks are 
normally required for review and 
circulation of the data prior to the 
meeting at which such data can be 
considered by the NIH Recombinant 
DNA Advisory Committee (RAC). 
Investigators are encouraged to publish 
their data on the construction, 
properties, and testing of proposed HV2 
systems prior to consideration of the 
system by the RAC and its 
subcommittee. More specific 
instructions concerning the type of data 
to be submitted to NIH for proposed EK2 
systems involving either plasmids or 
bacteriophage X in E. coli K-12 are 
available from ORDA. 
II— D— 2— b— (3). HV3 Systems. Putative 
HV3 systems must, as the first step in 
certification, be certified as HV2 
systems. Systems which meet the 
criteria given above under II-D-l-(c)-l, 
II— D— 1— (c)— 2, and II— D— 1— (c)— 3 will then 
be recommended for HV3 testing. Tests 
to evaluate various HV2 host-vector 
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