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Federal Register / Vol. 45, No. 147 / Tuesday, July 29, 1980 / Notices 
coli, experiments may be performed with any 
E. coli K-12 vector (e.g., conjugative plasmid). 
When a nonconjugative vector is used, the E. 
coli K-12 host may contain conjugation- 
proficient plasmids, either autonomous or 
intergrated, or generalized transducing 
phages.” 
II- G Amendment of Section III-A-2-a, 
Virsuses of Eukaryotes 
Substitute the term “an HVl host- 
vector” or “an HVl host and a vector 
certified for use in an HV2 system" in 
Section III-A-2-a-(l)-( a M 3 )-(b) and 
substitute the term "an HVl host- 
vector" for “an HVl host and a vector 
certified for use in an HV2 system, or 
P3 + HVl," in Sections III-A-2-a-(l)- 
(aH2Hc), III-A-2-a-(lHbHl)-(b), III- 
A-2-a-(lHb)-(2Hb), III-A-2-a-(2Ha)- 
(lHb), III-A-2-a-(2HaH2Hb), and 
III- A— 2— a— (2)— (c)— (2)— (b). In Table III, 
whenever any one of the following three 
terms appears, ("HVlCV[40]”; 
"HV1CVJ40] or P3 + HVl;” or 
"HV1CV[40] or P3 + HVl[38]"J. 
substitute “HVl.” 
II-H. Section III-A-3-a Is Amended as 
Follows 
III-A-3-a. Purified DN A Other Than 
Plasmids, Bacteriophages, and Other 
Viruses. The information of DNA 
recombinants from cellular DNAs that have 
been purified [41] and in which the absence 
of harmful sequences has been established 
[3] can be carried out under lower 
containment conditions than used for the 
corresponding shotgun experiment [42]. The 
containment may be decreased one step in 
physical containment [P4— *P3; P3— >P2; P2 
PI] while maintaining the biological 
containment specified for the shotgun 
experiement, or one step in biological 
containment [HV3— »HV2; HV2— »HVl] while 
maintaining Jhe specified physical 
containment. The institutional biosafety 
committee (1BC) must review such a 
reduction and the approval of the IBC must 
be secured before Such a reduction may be 
put into effect. IBC approval is sufficient for 
such a reduction except for any lowering of 
containment under Section III-A-3-a to 
levels below Pi + HVl, which requires prior 
NIH approval. (See Sections IV-D-l-c and 
IV- E-l-b-(3)(e).) 
//-/. Section III-A-3-b Is Amended as 
Follows 
III-A-3-b. Characaterized Clones of DNA 
Recombinants. When a cloned DNA 
recombinant has been rigorously 
characterized and the absence of harmful 
sequences has been established (3) 
experiments involving this recombinant DNA 
may be carried out under lower containment 
conditions. 
Institutional biosafety committees (IBCs) 
may give approval for a single-step reduction 
in physical or biological containment on 
receipt of evidence of characterization of a 
clone dervied from a shotgun experiment and 
its probable freedom from harmful genes. IBC 
approval is sufficient for such a reduction 
except for any lowering of containment under 
Section III-A-3-b to levels below PI + HVl, or 
reduction of containment levels by more than 
one step, which also requires prior NIH 
approval. (See Sections IV-D-l-c and IV-E-1- 
b-3-(e).) 
//-/ Amendment of Section III-B-3 
Section III-B-3, paragraph 2 is 
amended to read: 
In those cases where genetic exchange has 
not been demonstrated between two 
bacterial species A and B, neither of which is 
known to be pathogenic for man, animals or 
plants, recombinant DNA experiments 
involving only A and B can be conducted 
under P3 containment [2A]. Lower levels of 
physical containment may be assigned by 
NIH for specific donor-recipient 
combinations (See Section IV-E-l-b-2-(f)). 
Il-K. Amendment of Section III-C-3 of 
the Guidelines 
Delete paragraph four of Section III-C- 
3, which reads as follows: 
The CaMV strain used as a cloning vector 
shall be a mutant that lacks the aphid 
trasmission factor. 
II-L. Amendment of Section III-C-4 of 
the Guidelines 
Paragraph two of Section III-C-4 is 
amended by changing the end of the 
paragraph from “* * * work.” to “* * * 
work, and (iii) negative air pressure 
should be employed in the greenhouse 
or growth chamber when infectious 
agents are used which generate airborne 
propagules.” 
II-M. Amendment of Section III-C-7-b of 
the Guidelines 
Section IU-C-7-b is amended te read 
as follows: 
III- C-7-b. Transfer to Higher Plants. DNA 
from any nonprohibited source [Section I-D] 
which has been cloned and propagated in E. 
coli or S. cerevisiae under Pi conditions, may 
be transferred with the E. coli or S. 
cerevisiae vector used for cloning to any high 
plant organisms (angiosperms and 
Gymnosperms) and propagated under 
condition of physical containment 
comparable to PI and appropriate to the 
organism under study (2A). Intact plants or 
propagative plant parts may be grown under 
Pi conditions described under Section III-C-3. 
Containment must be modified to ensure that 
the spread of pollen, seed or other propagules 
is prevented. This can be accomplished by 
conversion to negative pressure in the growth 
cabinet or greenhouse or by physical 
entrapment by "bagging” of reproductive 
structures. Transfers to any other plant 
organisms will be considered on a case-by- 
case basis (45). 
II-N. Section IV-D-l-c-(3) Is Amended as 
Follows 
IV- D-l-c-(3). Certain reductions of 
containment levels for characterized or 
purified DNA preparations or clones (see 
Section III-A-3). 
II-O. Amendment of Section IV-D-3-b 
The note in Section IV-D-3-b is 
amended as follows: 
Note. — Some examples of work that might 
ordinarily proceed Without prior funding- 
agency approval are the initiation of a project 
at the PI or P2 level (other than the first 
project at the institution). Other examples are 
significant changes in hosts or vectors, in the 
donor species or the nature of the DNA 
segment selected, or in the physical location 
of the experiments. Still others are single-step 
reductions in containment level for (i) 
experiments with DNA recombinants from 
cellular DNAs that have been purified and 
are judged to be free of harmful sequences 
(See Section III-A-3-a) and for (ii) clones that 
have been characterized and judged to be - 
free of harmful sequences (see Section III-A- 
3-b). It should be clear, however, that the 
funding agency must be notified of IBC 
approvals even when prior agency approval 
is not required. See the Administrative 
Practices Supplement for further discussion. 
II-P. Addition of Section IV-E-l-b-2-(eJ 
A new Section IV-E-l-b-2-(e) is 
added as follows: 
IV-E-l-b-2-(e). Assigning containment 
levels for experiments with recombinant 
DNA from Class 3 organisms [1] and 
assigning containment levels for experiments 
which increase the host-range and virulence 
of plant pathogens beyond that which occurs 
by natural genetic exchange. 
II-Q. Addition of Section TV-E-l-b-2-(f) 
A new Section IV-E-l-b-2-(f) is 
added a follows: 
IV-E-l-b-2-(f). Assigning containment 
levels for experiments in which both donor 
and recipient are non-pathogenic prokaryotes 
(See Section III-B-3). 
ll-R. Amendment of Footnote 2, Section 
V 
Section V, footnote 2, is amended to 
read: 
2. For experiments using Vesicular 
Stomatitis virus (VSV), contact the NIH 
Office of Recombinant DNA Activities. 
II-S. Amendment of Footnote 36, Section 
V 
The second sentence of footnote 38, 
Section V is amended to read: 
(As noted in the Prohbition Section, the use 
of viruses classified [1] as Class 4 or 5 is 
prohibited.) 
II-T. Status of Variola and Whitepox 
Viruses 
A new footnote is added to Appendix 
B of the Guidelines as follows: 
* * * All activities, including storage of 
variola and whitepox are restricted to the 
single national facility (World Health 
Organization (WHO) Collaborating Center 
[147] 
