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Federal Register / Vol. 47, No. 77 / Wednesday. April 21, 1982 / Notices 
DEPARTMENT OF HEALTH AND 
HUMAN SERVICES 
Quidellnee for Research Involving 
Recombinant DNA Molecules; April 
1982 
Table of CootenU 
L Scope of Ihe Cuidelinea 
l-A — Puipoee 
l-B— Definition of Recombinant DNA 
Molecule* 
l-C — General Applicability (*ee IV-B) 
1-0— General Oeflnitiona (»ee IV-C) 
n. Containment 
0- A — Standard Practice* and Training 
1- B— Phy*ical Containment Level* 
n-B-l- Pi Level 
ll-B-l-a — Laboratory Practice* 
Il-B-l-b— Containment Equipment 
H-B-1-C — Special Laboratory Deaign 
ll-B-2— P2 Level 
I1-B-2-* — Laboratory Practice* 
O-B-2-b— Containment Equipment 
U-B-2-C — Special Laboratory De*ign 
U-8-3— P3 Level 
U-B-3-a — Laboratory Practice* 
Il-B-3-b— Containment Equipment 
D-8-3-C— Special Laboratory De*ign 
D B 4 P* Level 
D' B 4 a — Laboratory Practice* 
D -B -4 ' b— Containment Equipment 
D B 4 c — Special Laboratory Denign 
n-C — Shipment 
B-C — Biological Containment 
D-O-1— Level* of Biological Containment 
D-O-l-a— HVl 
D-O-l-b— HV2 
D-O-2 — Certification of Hoat-Vector 
Sy*tem* 
Il-O-2-a — Reeponaibility 
D-O-2-b— Data To Be Submitted for 
Certification 
ll-O-S— Diatributlon of Certified Hoet- 
Vector* 
01. Containment Guideline* for Covered 
Experiment* 
Ql-A — Experiment* That Require RAC 
Review and NIH and IBC Approval 
Before Initiabon 
m-B— Experiment* that Require IBC 
Approval Before Initiation 
m-^l — Experiment* U*ing CDC Qa** 2. 
Qaa* 3. Qaaa 4. or Qa** 5 Agent* a* 
Hoat-Vector Syatem* 
ID-B-2 — Experiment* in which DNA from 
CDC Qaa* 2. Claa* 3. Qa** 4. or Qa** S 
Agent* I* Cloned in Nonpathogenic 
Prokaryotic or Lower Eukaryotic Ho*t- 
Vector Syatem* 
IU-B-3 — Experiment* Involving the U*e of 
Infection* Animal or Plant Viruae* or 
Defective Animal or Plant Viruae* in the 
Preaence of Helper Viru* in Tlaaue 
Culture Syatem* 
m-B-4 — Recombinant DNA Experiment* 
Involving Whole Animal* and Plant* 
IH-B-5— Experiment* Involving More than 
10 Liter* of Culture 
m-C — Experiment* that Require IBC 
Notice Simultaneoualy with Initiation of 
Experiment* 
m-D— Exempt Experiment* 
rV. Rule* and Reaponaibilitie* 
tV-A— Policy 
rV-B — General Applicability 
rV-C — General Definitions 
IV-D — Responsibilities of Ihe Institution 
IV-D-1 — (General) 
rV-D-2 — Membership and Procedures of 
the IBC 
lV-D-3 — Functions of the IBC 
IV-D-4 — Biological Safety Officer 
IV-D-5— Principal Investigator 
IV-D-5-a — PI — General 
IV-D-5-b — Submissions by Ihe PI to NIH 
fV-D-5-c — Submissions by the PI to the 
IBC 
IV-D-5-d — PI Responsibilities Prior to 
Initiating Research 
IV-D-5-e — PI Reaponaibilitie* During the 
Conduct of Ihe Research 
IV-E — Responsibilities of NIH 
IV-E-1 — Director 
IV-E-l-a — General Responsibilities of the 
Director, NIH 
rV-E-1-b— Specific Reaponaibilitie* of the 
Director, NIH 
IV-E-2 — Recombinant DNA Advisory 
Committee 
IV-E-3 — The Office of Recombinant DNA 
ArtivIlie* 
IV-E-4 — Other NIH Component* 
IV-P — Compliance 
V. Footnote* and Reference* 
VI. Voluntary Compliance 
Vl-A — Basic Policy 
Vl-B— IBC Approval 
Vl-C— Certification of Host-Vector Syatem. 
Vl-D— Requaata for Exemption* and 
Approvals 
Vl-E— Protection of Proprietary Data 
Appendix A — Fjiemptions Under llI-D-4 
Appendix B— Qaasification of 
Microorganisms on the Basis of Hazard 
Appendix C— Exemption* Under III-D-5 
Appendix D — Action* Taken Under the 
Guideline* 
Appendix E — Certified Host-Vector Systems 
Appendix F — Containment Conditions for 
Cloning of Gene* Coding for Ihe 
Biosynthesis of Toxins for Vertebrates 
I. Scope of the Guidelines 
I-A. Purpose. The purpose of these 
Guidelines is to specify practices for 
constructing and handling (i) 
recombinant DNA molecules and (ii) 
organisms and viruses containing 
recombinant DNA molecules. 
I-B. Definition of Recombinant DNA 
Molecules. In the context of these 
Guidelines, recombinant DNA molecules 
are defined as 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 that result from the 
replication of those described in (i) 
above. 
Synthetic DNA segments likely to 
yield a potentially harmful 
polynucleotide or polypeptide (e.g.. a 
toxin or a pharmacologically active 
agent) shall be considered as equivalent 
to their natural DNA counterpart. If the 
synthetic DNA segment is not expressed 
in vivo as a biologically active 
polynucleotide or polypeptide product, it 
is exempt from the Guidelines. 
I-C. General Applicability. See 
Section IV-B. 
I-D General Definitions. See Section 
IV-C. 
II. Containment 
Effective biological safety programs 
have been operative in a variety of 
laboratories for many years. 
Considerable information, therefore, 
already exists for the design of physical 
containment facilities and the selection 
of laboratory procedures applicable to 
organisms carrying recombinant DNAs 
(6-19). The existing programs rely upon 
mechanisms that, for convenience, can 
be divided into two categories; (i) A set 
of standard practices that are generally 
used in microbiological laboratories, 
and (ii) special procedures, equipment, 
and laboratory installations that provide 
physical barriers which are applied in 
varying degrees according to the 
estimated biohazard. 
Experiments on recombinant DNAs. 
by their very nature, lend themselves to 
a third containment mechanism — 
namely, the application of highly 
specific biological barriers. In fact, 
natural barriers do exist which limit 
either (i) the infectlvity of a vector, or 
vehicle, (plasmid or virus) for specific 
hosts or (ii) its dissemination and 
survival in the environment. The vectors 
that provide the means for replication of 
the recombinant DNAs and/or the host 
cells in which they replicate can be 
genetically designed to decrease by 
many orders of magnitude the 
probability of dissemination of 
recombinant DNAs outside the 
laboratory. 
As these three means of containment 
are complementary, different levels of 
containment appropriate for 
experiments with different recombinants 
can be established by applying various 
combinations of the physical and 
biological barriers along with a constant 
use of the standard practices. We 
consider these categories of 
containment separately here in order 
that such combinations can be 
conveniently expressed in the 
Guidelines. 
In constructing these Guidelines, it 
was necessary to define boundary 
conditions for the different levels of 
physical and biological containment and 
for the classes of experiments to which 
they apply. We recognize that these 
definitions do not take into acount all 
existing and anticipated information on 
special procedures that will allow 
particular experiments to be carried out 
(413] 
