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Federal Register / Vol. 44, No. 232 / Friday, November 30, 1979 / Notices 
purified cDNA copies of viral mRNA. Strand RNA Viruses Belonging to 
[37] Presently Classified Viral Families. [36] 
III-A-2-a-(2)-(c)-{2). Other Plus- 
Table III .— Recommended Containment lor Cloning of Viral ON A or cDNA in Certain HV1 and HV2 Systems 
Specified in Appendix D 
[See text for full details] 
Type of viral DNA segment to be cloned 
Virus class Subgenom»cl38] Genomic* 
cONA from viral 
mRNAC37] 
Segment 
Non transforming containing an Nonsegmented Segmented 
segment entire genome genome 
transforming 
gene 
DNA 
Nontransforming 
viruses 
AAV, MVM, Mouse Adeno (Strain FL)... PI + HV1 
Plant Viruses .. .. P.1+HV1 
Hepatitis B Pi + HVl [38].... 
Ottier .. PI + HV1[38]..„ 
Transforming Viruses 
P1+HV1. 
PI +HV1 
P2 + HV2 Of 
P3 + HV1. 
PI +HV1CVC40] 
Herpes Satnwi. K Ateles and PI +HV1 [38].... P2 + HV1CV[40] P2 + HV2 or 
EBV[39J. or P3 + HV1. 
P3 + HVU38], 
Other PI + HV1[38].... P2 + HV1CV[40] P2 + HV1CV[40] 
or P3 + HV1. or P3 + HV1. 
PI +HV1 
PI +HV1 
P2 + HV1CVt40] 
or P3 + HV1 
PI + HVl 
P2 + HV1CV[40] 
or P3 + HV1 
P2 4 HV1CV[40] 
or P3 + HV1 
RNA 
Retro viruses 
Gibbon Ape. WooHy Monkey FeLV and 
FeSV[39]. 
Negative-Strand RNA 
Plus-Strand RNA 
Types 1 and 2 Sabin Polio, 17D Yellow 
Fever Vaccine Strains 
Other .. 
Double-Stranded RNA 
Plant Viruses -fViroids 
Intracellular Viral DNA 
PI + HV1 [38].... P2 + HV1CVr40] P2 + HV2 or 
or P3 + HV1. 
P3-I-HV1 [38], 
P1+HV1[38]...*P2 + HV1CV[40] P2 + HV1CV[401 
Of P3 + HV1. or P3 + HV1. 
P1+HV1 PI +HV1 PI +HV1 .. 
Pi + HVl PI + HVl ..... 
PI + HVU38].... .. P2-fHVlCV[40] 
or P3 + HV1. 
P1+HV1 P1+HV1 
PI+HVt - - Pt + HVI. PI +HV1 
See text See text See text 
P2 + HV2 or 
P3 + HV1 
P2 + HV1CV[40] 
or P3 + HV1 
Pi +HV1 
Pi + HVl 
P2 + HV1CV[40] 
Of P3 + HV1 
P1+HV1 
PI + HVl 
*See exception given at asterisk at end of Appendix D. 
III-A-2-a-(2)-(c)-(2H°)- Pi physical 
containment + an HVl host-vector shall 
be used for DNA recombinants 
produced with purified subgenomic 
cDNA segments.[38] 
III- A-2-a-( 2 )-( c )-(2)-{6 ) . P2 physical 
containment + and HVl host and a 
vector certified for use in an HV2 
system, or P3 + HVl, shall be used for 
DNA recombinants produced with (i) 
cDNA copies of the whole genome, or 
(ii) purified cDNA copies of viral 
mRNA. [37] 
III-A-2-a-(2)-(d). Double-Stranded 
Segmented RNA Viruses. Pi physical 
containment + an HVl host-vector shall 
be used for DNA recombinants 
produced with (i) mixtures of 
subgenomic cDNA segments, [ii] a 
specific subgenomic cDNA segment, or 
(iii) purified cDNA copies of viral 
mRNA.[37] 
III-A-2-a-(2)-(e). RNA Plant Viruses 
and Plant Viroids. PI physical 
containment + an HVl host-vector shall 
be used for DNA recombinants 
produced with (i) cDNA copies of the 
whole viral genome, (ii) subgenomic 
cDNA segments, or (iii) purified cDNA 
copies of viral mRNA. [37] 
III-A-2-a— (3). Intracellular Viral 
DNA. Physical and biological 
containment specified for shotgun 
experiments with eukaryotic cellular 
DNA [see Section III— A— (1)— (a)] shall be 
used for DNA recombinants produced 
with integrated viral DNA or viral 
genomes present in infected cells. 
III-A-2-b. Eukaryotic Organelle 
DNAs. P2 physical containment + an 
HVl host-vector, or PI + HV2, for 
mitochondrial or chloroplast DNA from 
eukaryotes when the organelle DNA has 
been obtained from isolated organelles. 
[ 206 ] 
Otherwise, the conditions given for 
shotgun experiments apply. 
IIl-A-2-c. Prokaryotic-Plasmid and 
Phage DNAs. The containment levels 
required for shotgun experiments with 
DNA from prokaryotes apply to then - 
plasmids or phages (See Section III— A— 
1— b.) 
III-A-3. Lowering of Containment 
Levels for Characterized or Purified 
DNA Preparations and Clones. Many of 
the risks which might conceivably arise 
from some types of recombinant DNA 
experiments, particularly shotgun 
experiments, would result from the 
inadvertent cloning of a harmful 
sequence. Therefore, in cases where the 
risk of inadvertently cloning the 
“wrong” DNA is reduced by prioi 
enrichment for the desired piece, or in 
which a clone made from a random 
assortment of DNAs has been purified 
and the absence of harmful sequences 
established, the containment conditions 
for further work may be reduced. The 
following section outlines the 
mechanisms for such reductions. 
III-A-3-a. Purified DNA Other than 
Plasmids, Bacteriophages, and Other 
Viruses. The formation 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 ^4— ►P3; P3— *P2; P2— »Pl) 
while maintaining the biological 
containment specified for the shotgun 
experiment, or one step in biological 
containment (HV3— ►HV2; HV2— ►HVl) 
while maintaining the specified physical 
containment. The institutional biosafety 
committee (IBC) must review such a 
reduction and the approval of the IBC 
and of the NIH must be secured before 
such a reduction may be put into effect. 
III-A-3-b. Characterized 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, 
with the prior approval of the IBC and of 
NIH. 
Ill— B. Experiments with Prokaryotic 
Host- Vectors Other Than E. coli K-12 
III— B— 1. HVl and HV2 Systems. 
Certain certified HVl and HV2 host- 
vector systems appear in Appendix D. 
The containment levels for these 
systems are given in the subsections of 
Section III— A. Other systems in the 
