Federal Register / Vol. 46, No. 233 / Friday, December 4, 1981 / Notices 
59375 
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 what 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. 
lI-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 hosWector 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 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. 
Il-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-l-(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 selectd by 
NIH. These contractors will repeat tests 
performed by individuals proposing the 
HV2 system and, in 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 distributing 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 should 
first consult Part I, where exempt 
experiments are listed. 
Where recommended physical 
containment levels applicable to non- 
recombinant DNA experiments exist for 
either the host or the vector,* 
recombinant DNA experiments should 
be carried out at containment levels at 
least as high as those recommended for 
non-recombinant DNA experiments. If 
there is clear evidence that the donor 
DNA will significantly change the 
pathogenicity of the host, the 
containment level appropriate to the 
anticipated change will be applied. 
Otherwise, all experiments may be 
carried out imder conditions of PI or PI- 
TS physical containment. 
No experiments should be performed 
which involve: 
(a) Deliberate transfer of a drug 
resistance trait to microorganisms that 
are not known to acquire it naturally, if 
such acquisition could compromise the 
use of a drug to control disease agents in 
human or veterinary medicine or 
agriculture, 
(b) Deliberate formation of 
recombinant DNAs containing genes for 
the biosynthesis of toxins lethal for 
vertebrates at an UOm of less than.lOO 
nanograms per kilogram body weight 
(e.g., the botulinum toxins, tetanus toxin, 
diphtheria toxin. Shigella dysenteriae 
neurotoxin). Guidelines for the cloning 
of DNAs containing genes coding for the 
biosynthesis of toxins which are lethal 
to vertebrates at 100 nanograms to 100 
micrograms per kilogram body weight 
are specified in Appendix G. 
IV. Roles and Responsibilities 
IV-A. [Deleted] 
IV-B. [Deleted] 
IV-C. General Definitions. The 
following terms are defined as follows: 
IV-C-1. "DNA” means 
deoxyribonucleic acid. 
IV-C-2. "Recombinant DNA” or 
"recombinant DNA molecules” means 
either (i) molecules which are 
constructed outside living cells by 
joining natural or synthetic DNA 
segments to DNA molecules that can 
replicate in a living cell, or (ii) DNA 
molecules which result from the 
replication of a molecule described in (i) 
above. 
IV-C-3. [Deleted] 
IV-C-4. "Institution” means any 
public or private entity (including 
Federal, State, and local government 
agencies). 
IV-C-5. [Deleted] 
IV-C-6. "NIH Office of Recombinant 
DNA Activities” or "ORDA” means the 
office within NIH with responsibility for 
* Such as those specified by the CDC Guidelines, 
or the USDA Quarantine Regulations. 
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