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Federal Register / Vol. 47, No. 77 / Wednesday, April 21, 1982 / Notices 
(3Hd).) 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system is rescinded, NIH will instruct 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 fur 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. 
n-D-2-b-(l). 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 what sort of information is 
exchanged; and any relevant 
information on its pathogenicity or 
toxity. (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 
stirvive or transmit genetic information, 
(iii) A general description of the range of 
experiments contemplated, with 
emphasis on the need for developing 
such an HlL 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 
tranformation. (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 FJC2 
systems involving either palsmids or 
bacteriophage in E coli K-12 are 
available from ORDA. 
ll-D-3. Distribution of Certified Host- 
Vectors. Certified HV2 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 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 
involving recombinant DNA. These 
experiments have been divided into four 
classes; 
Ill-A. Experiments which require 
specific RAC review and NIH and IBC 
approval before initiation of the 
experiment; 
III-B. Experiments which require IBC 
approval before initiation of the 
experiment; 
III-C. Experiments which require IBC 
notification at the time of initiation of 
the experiment; 
III-D. Experiments which are exempt 
from the procedures of the Guidelines. 
IF AN EXPERIMENT FALLS INTO 
BOTH CLASS III-A AND ONE OF THE 
OTHER CLASSES. THE RULES 
PERTAINING TO CLASS III-A MUST 
BE FOLLOWED. If an experiment falls 
into class III-D and into either class III- 
B or IIl-C as well, it can be considered 
exempt from the requirements of the 
Guidelines.' 
Changes in containment levels from 
those specified here may not be 
instituted without the express approval 
of the Director, NIH. (See Sections IV- 
E-l-b-(l). IV-E-l-b-(2). and 
subsections.) 
III-A. Experiments that Require RAC 
Review and NIH and IBC Approval 
Before Initiation. Experiments in this 
category cannot be initiated without 
submission of relevant information on 
the proposed experiment to NIH, the 
publication of the proposal in the 
Federal Register for thirty days of 
comment, review by the RAC, and 
specific approval by NIH. The 
containment conditions for such 
experiments will be recommended by 
RAC and set by NIH at the time of 
approval. Such experiments also require 
the approval of the IBC before initiation. 
Specific experiments already approved 
in this section and the appropriate 
containment conditions are listed in 
Appendices D and F. 
IIl-A-1. Deliberate formation of 
recombinant DNAs containing genes for 
the biosynthesis of toxins lethal for 
vertebrates at an LDm of less than 100 
nanograms per kilogram body weight 
(e.g., the bottilinum toxins, tetanus toxin, 
diphtheria toxin. Shigella dysenteriae 
neurotoxin). Specific approval has been 
given for the cloning in E. coli K-12 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 
Containment levels for these 
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