60114 
NOTICES 
II-D-l-a-(2). Other Prokaryotes. 
Hosts and vectors shall be, at a mini- 
mum, comparable in containment to E. 
Col K-12 with a non conjugative plas- 
mid or bacteriophage vector. The data 
to be considered and mechanism for 
approval of such HV1 systems are de- 
scribed below (Section H-D-2). 
II-D-l-b. HV2 These axe host-vector 
systems shown to provide a high level 
of biological containment as demon- 
strated by data from suitable tests per- 
formed in the laboratory. Escape of 
the recombinant DNA either via sur- 
vival of the organisms or via transmis- 
sion of recombinant DNA to other or- 
ganisms should be less than 14 »• under 
specified conditions. Specific systems: 
II-D-l-tHl). For EK2 host-vector 
systems in which the vector is a plas- 
mid, no more than one in 10" host cells 
should be able to perpetuate a cloned 
DNA fragment under the specified 
nonpermissive laboratory conditions 
designed to represent the natural envi- 
ronment, either by survival of the 
original host or as a consequence of 
transmission of the cloned DNA frag- 
ment. 
II-D-l-b-(2). For EK2 host-vector 
systems in which the vector is a phage, 
no more than one in 10" phage parti- 
cles should be able to perpetuate a 
cloned DNA fragment under the speci- 
fied nonpermissive laboratory condi- 
tions 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 nat- 
ural environments and transferring a 
cloned DNA fragment to other hosts 
(or their resident prophages). 
n-D-l-c. HV3. These are host-vector 
systems in which: 
II-D-l-c-(l). All HV2 criteria are 
met. 
II-D-l-c-(2). The vector is depend- 
ent on its propagation host or is 
highly defective in mobilizability. Re- 
version to host-independence must be 
less than yio* per vector genome per 
generation. 
II-D-l-c-(3). No markers conferring 
resistance to antibiotics commonly 
used clinically or in agriculture are 
carried by the vector, unless expres- 
sion of such markers is dependent on 
the propagating host or on unique lab- 
oratory-controlled conditions or is 
blocked by the inserted DNA. 
n-D-l-c-(4). The specified contain- 
ment shown by laboratory tests has 
been independently confirmed by 
specified tests in animals, including 
primates, and in other relevant envi- 
ronments. 
II-D-l-c-(5). The relevant genotypic 
and phenotypic traits have been inde- 
pendently confirmed. 
II-D-2. Certification of Host-Vector 
Systems. 
H-D-2-a. Responsibility. HV1 sys- 
tems 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 ap- 
plication to the Office of Recombinant 
DNA Activities (ORDA), National In- 
stitutes of Health, Bethesda, Mary- 
land 20014. 
Host-vector systems that are pro- 
posed for certification will be reviewed 
by the NIH Recombinant DNA Adviso- 
ry Committee (RAC). (See Section IV- 
E-l-b-GMO.) 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 mem- 
bers 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 availa- 
ble 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 mini- 
mal or no consequence to the proper- 
ties relevant to containment may be 
certified by the Director, NIH without 
review by the RAC. (See Section IV-E- 
l-b-(3)-(h).) 
When new host-vector systems are 
certified, notice of the certification 
will be sent by ORDA to the applicant 
and to all EBCs and will be published 
in the Recombinant DNA Technical 
Bulletin. Copies of a list of all current- 
ly certified host-vector systems may be 
obtained from ORDA at any time. 
The Director, NTH may at any time 
rescind the certification of any host- 
vector system. (See Section TV-E-l-b- 
(3 Mi).) 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 contain- 
ment level unless NIH determines that 
the already constructed clones Incor- 
porate adequate biological contain- 
ment. 
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 Di- 
rector, NTH. If modifications are 
minor, it may only be necessary for 
the investigator to submit data show- 
ing that the modifications have either 
improved or not impaired the major 
phenotypic traits on which the con- 
tainment of the system depends. Sub- 
stantial modifications of a certified 
system require the submission of opm- 
plete testing data. 
II-D-2-b. Data To Be Submitted for 
Certification. 
n-D-2-b-(l). HV1 Systems Other 
than E. Coli K-12. The following types 
of data shall be submitted, modified as 
appropriate for the particular system I 
under consideration: (i)A description 
of the organism and vector; the 
strain’s natural habitat and growth re- 
quirements; its physiological proper- 
ties, particularly those related to its 
reproduction and survival and the 
mechanisms by which it exchanges ge- 
netic information; the range of organ- i 
isms with which this organism normal- / 
ly exchanges genetic information and | 
what sort of information is exchanged; 
and any relevant information on its 
pathogenicity or toxicity, (ii) A de- 
scription of the history of the particu- 
lar strains and vectors to be used, in- 
cluding data on any mutations which 
render this organism less able to sur- | 
vive or transmit genetic information. , 
(iii) A general description of the range 
of experiments contemplated, with : 
emphasis on the need for developing r 
such an HV1 system. • 
II-D-2-b-(2). HV2 Systems. Investi- 
gators planning to request HV2 certifi- (, 
cation for host-vector systems can s 
obtain instructions from ORDA con- 
cerning data to be submitted [33A, |j 
33B1. In general, the following types y 
of data are required: (i) Description of j 
construction steps, with indication of e 
source, properties, and manner of in- 
troduction of genetic traits, (ii) Quan- 
titative data on the stability of genetic 
traits that contribute to the contain- 
ment of the system, (iii) Data on the 
survival of the host-vector system 
under non-permissive laboratory con- 
ditions designed to represent the rele- h 
vant natural environment, (iv) Data on < 
transmissibility of the vector and/or a L 
cloned DNA fragment under both per- 
missive 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 t 
DNA isolation, and ease of transfec- 
tion 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 labora- 
tory animals (e.g., rodents). Such in y 
vivo data may be required to confirm J 
the validity of predicting in vivo sur- J 
vival on the basis of in vitro experi- 
ments. 
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 1 
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- 
FEDERAl REGISTER, VOL. 43, NO. 247— FRIDAY, DECEMBER 22, 1978 
[ 36 ] 
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