33076 
NOTICES 
be used unless they have been certi- 
fied by the NIH. Application for certi- 
fication of a host-vector system is 
made by written application to the 
Office of Recombinant DNA Activities 
(ORDAV National Institutes of 
Health. Bethesda. Md. 20014. 
When appropriate, the proposed 
host-vector system will be reviewed by 
the NIH Recombinant DNA Advisory 
Committee (RAC). This may first in- 
volve review of the data on construc- 
tion. properties, and testing of the pro- 
posed host-vector system by a subcom- 
mittee composed of one or more mem- 
bers of the RAC and other individuals 
chosen because of their expertise in 
evaluating such data. The committee 
will then evaluate the report of the 
subcommittee and any other available 
information at a regular meeting. 
When new host-vector systems are 
certified, notice of the certification 
will be sent to the applicant and to all 
IBCs and will be published in the "re- 
combinant DNA Technical Bulletin.'* 
Copies of a list of all currently certi- 
fied host-vector systems may be ob- 
tained from ORDA at any time. 
NIH may at any time rescind the 
certification of any host-vector 
system. If cert if cation of a host-vector 
system is rescinded, investigators may 
be asked to transfer cloned DNA into a 
different system. 
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 NIH. If 
modifications are minor, it may only 
be necessary for the investigator to 
submit data showing that the modifi- 
cations have either Improved or not 
Impaired the major phenotypic traits 
on which the containment of the 
system depends. More substantial 
modifications of a certified system 
may necessitate submission of com- 
plete testing data. 
II-D-2-b Data to be Submitted for 
Certification 
II-D2-b-< 1). HVl System* Other than 
E. Coti K-IZ. The follow-tng types of 
data should be submitted, modified as 
appropriate for the particular system 
under consideration: (1) A description 
of the organism and vector, the 
strain’s natural habitat and growth re- 
quirements: the range of organisms 
with which this organism normally ex- 
changes genetic Information and what 
sort of information is exchanged: any 
relevant information on Its pathogeni- 
city 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 trans- 
mit genetic Information, (ill) A general 
description of the range of experi- 
ments contemplated, with emphasis on 
the need for developing such an HVl 
system. 
I- D-2-b-<2>. HVZ Systems. Investiga- 
tors planning to request HV2 certifica- 
tion for host-vector systems can obtain 
instructions from ORDA concerning 
data to be submitted. 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. <li) Quantitative data on the 
stability of genetic traits that contrib- 
ute to the containment of the system, 
(iii) Data on the survival of the host- 
vector system under non-permissive 
laboratory conditions designed to rep- 
resent the relevant natural environ- 
ment. (iv) Data on transmissibility of 
the vector and/or a cloned DNA frag- 
ment under both permissive and non- 
permissive conditions, (v) Data on all 
other properties of the system which 
affect containment and utility, includ- 
ing Information on yields of phage or 
plasmid molecules, ease of DNA isola- 
tion. and ea..e of transfection or trans- 
formation. (vi) In some cases, the in- 
vestigator may be asked to submit 
data on survival and vector transmissi- 
bility from experiments In which the 
host-vector is fed to laboratory ani- 
mals <e.g.. rodents). Such in mo data 
may be required to confirm the valid- 
ity of predicting in tiro survival on 
the basis of in vitro experiments. 
Data must be submitted in writing to 
ORDA. Ten to twelve weeks are nor- 
mally required for review and circula- 
tion of the data prior to the meeting 
at which such data can be considered 
by the NIH Recombinant DNA Adviso- 
ry Committee (RAC). Investigators are 
encouraged to publish their data on 
the construction, properties, and test- 
ing of proposed HV2 systems prior to 
consideration of the system by the 
RAC and Its subcommittee. More spe- 
cific instructions concerning the type 
of data to be submitted to NIH for 
proposed EK2 systems Involving either 
plasmids or bacteriophage 0 in E. coti 
K-12 are available from ORDA 
II- D-2-b-<3). HVZ Systems. Putative 
HV3 systems must, as the first step in 
certification, be certified as HV2 sys- 
tems. 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-sector 
systems for HV3 certification will be 
performed by contractors selected by 
NIH. These contractors will repeat 
tests performed by Individuals propos- 
ing 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 evaluat- 
ed. Tests on survival and transmissibil- 
ity in and on animals, including pri- 
mates. will be performed, as well as 
tests on survival in certain specified 
natural environmnets. 
II- D-3. Distribution of Certified 
Host-Vectors. Certified HY2 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 publica- 
tion of notices in appropriate journals. 
Plasmid vectors will be provided in a 
suitable host strain, and phage vectors 
will be distributed as small-volume ly- 
sates. 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 con- 
tractor. prior to distribution, for verifi- 
cation 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) enumer- 
ate and describe the tests to be per- 
formed by the user in order to verify 
important phenotypic traits: (iii) 
remind the user that any modification 
of the system necessitates independ- 
ent approval of the system by the NIH 
on recommendation of the RAC; and 
<lv) remind the user of responsibility 
for notifying ORDA of any discrepan- 
cies 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 cov- 
ered by the guidelines. The reader 
must first consult part I. where list- 
ings are given of prohibited and 
exempt experiments. 
Containment guidelines for permissi- 
ble experiments are given in part III. 
Changes in these levels for specific ex- 
periments (or the assignment of levels 
to experiments not explicitly consid- 
ered here) may be expressly approved 
by the Director. NIH. on the recomme- 
datlon of the Recombinant DNA Advi- 
sory Committee (RAC). 
III- A. Classification of Experiments 
Using the E. coti K-12 Host-Vector Sys- 
tems. Most recombinant DNA experi- 
ments currently being done employ E. 
coti K-12 host-vector systems. These 
are the systems lor which we have the 
most experience and knowledge (i) re- 
garding the effectiveness of biological 
containment provided by existing 
hosts and vectors, and (ii) necessary 
for the construction of more effective 
biological barriers. We therefore con- 
sider DNA recombinants in E. coti K- 
12 before proceeding to other host- 
vector systems. The levels of biological 
containment for E. coti K-12 systems 
FEDERAL REGISTER, VOL 43. MO. 144 — FRIDAY, JUIY tt, 1978 
[ 37 ] 
