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Federal Register / Vol. 46, No. 233 / Friday. December 4, 1981 / Notices 
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 
properities 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 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 in E. coli K-12 are 
available from ORDA. 
U-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 critera 
given above under II-D-l-(c)-l, II-D-1- 
(c)-2, and II-D-l-(c)-3 will then be 
recommended for HV3 testing. Tests to 
evaluate various HV2 host-vector 
systems for HV3 certification will be 
performed by contractors selected by 
NIH. These contractors will repeat tests 
performed by individuals proposing the 
HV2 system and, m addition, will 
conduct more extensive tests on 
conditions likely to be encountered in 
nature. The genotypic and phenotypic 
traits of HV2 systems will be evaluated. 
Tests on survival and transmissibility in 
and on animals, including primates, will 
be performed, as well as tests on 
survival in certain specified natural 
environments. 
II-D-3. Distribution of Certified Host- 
Vectors. Certified HV2 and HV3 host- 
vector systems (plus appropriate control 
strains) must be obtained from the NIH 
or its designees, one of whom will be the 
investigator who developed the system. 
NIH shall announce the availability of 
the system by publication of notices in 
appropriate journals. 
Plasmid vectors will be provided in a 
suitable host strain, and phage vectors 
will be distributed as small-volume 
lysates. If NIH propagates any of the 
host strains or phage, a sample will be 
sent to the investigator who developed 
the system or to an appropriate 
contractor, prior to distribution, for 
verification that the material is free from 
contamination and unchanged in 
phenotypic properties. 
In disteibuting the certified HV2 and 
HV3 host-vector systems, NIH or its 
designee will (i) send out a complete 
description of the system; (ii) enumerate 
and describe the tests to be performed 
by the user in order to verify important 
phenotypic traits: (iii) remind the user 
that any modification of the system 
necessitates independent approval of 
the system by the NIH; and (iv) remind 
the user of responsibility for notifying 
ORDA of any discrepancies with the 
reported properties or any problems in 
the safe use of the system. 
NIH may also distribute certified HVl 
host-vector systems. 
III. Containment Guidelines for Covered 
Experiments 
Part III discusses experiments covered 
by the Guidelines. The reader must first 
consult Part I, where listings are given of 
prohibited and exempt experiments. 
Containment guidelines for 
permissible experiments are given in 
Part III. For these experiments no 
registration with the National Institutes 
of Health (NIH) is necessary. However, 
for these experiments, prior to their 
initiation, investigators must submit to 
their Institutional Biosafety Committee 
(IBC) a registration document that 
contains a description of (a) the 
source(s) of DNA, (b) the nature of the 
inserted DNA sequences, (c) the hosts 
and vectors to be used, (d) whether a 
deliberate attempt will be made to 
obtain expression of a foreign gene in 
the cloning vehicle and if so, what 
protein, and (e) the containment 
conditions specified by these 
Guidelines. This registration document 
must be dated and signed by the 
investigator and filed only with the local 
IBC. The IBC shall review all such 
proposals: IBC review prior to initiation 
of the experiment is not required for 
experiments described in Section III-O. 
Prior IBC review is required for all other 
experiments described in the 
subsections of Part III, including III-O-l, 
III-0-2, etc. 
Changes from the levels specified in 
Part III for specific experiments (or the 
assignment of levels to experiments not 
explicitly considered here) may not be 
instituted without the express approval 
of the Director, NIH. (See Sections IV- 
E-l-b-(l)-(a), IV-E-l-b-(l)-(b), IV-E-1- 
b-(2)-(b). IV-E-l-b-(2)-(c), IV-E-l-b- 
(3)-(b).) 
In the classification of containment 
criteria for different kinds of 
recombinant DNAs, the stated levels of 
physical and biological containment are 
minimal for the experiments designated. 
The use of higher levels of biological 
containment (HV3>HV2>HVl) is 
encouraged if they are available and 
equally appropriate for the purposes of 
the experiment. 
When the reader finds 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. 
III-O. Classification of Experiments 
Using Certain Host-Vector Systems. 
Experiments listed in Appendix H may 
be performed at Pi physical 
containment. For these experiments IBC 
review prior to initiation of the 
experiment is not required. 
III-O-l. Experiments Involving Class 
3 Organisms. Experiments involving 
recombinant DNA from Class 3 
organisms [1] or from cells known to be 
infected with these agents may be 
conducted at P3 containment in E. coli 
K-12 EKl hosts (see Appendix C). 
Containment levels for all other 
experiments with Class 3 organisms or 
with recombinant DNA which increases 
the virulence and host range of a plant 
pathogen beyond that which occurs by 
natural genetic exchange will be 
determined by NIH. (See Section IV-E- 
l_b-2-(e)). 
ni-0-2. Experiments involving 
Nonpathogenic Prokaryotic and Lower 
Eukaryotic Host-Vector Systems. DNA 
from any species nonpathogenic for 
man, animals, or plants may be cloned 
into lower eukaryotes nonpathogenic for 
man, animals, or plants at the P3 level of 
[295] 
