Federal Register / Vol. 46, No. 126 / Wednesday, July 1. 1981 / Notices 
34487 
Neurvapora crassa ligated to a mobilizable 
plasmid 
34. E. coU K-12 strain DF214 (or derivatives 
thereof) and plasmid vectors (e.g.. pBR322. 
pBR325) may be used to clone rat cDNA 
under P2 conditions. After the clone of 
interest has been puriHed. it may be worked 
with the under Pi containrpent. 
Appendix F. — Certifled Host-Vector Systems 
While many experiments using £. coli K- 
12. Saccharomyces cerevisiae and Bacillus 
BubtiHs are currently exempt from the 
Guidelines under Exemption l-E-5. some 
derivatives of these host-vector systems were 
previously classified as HVl or HV2. A 
listing of those systems follows. 
HVl — The following plasmids are accepted 
as the vector components of certiFied B. 
subtUis HVl systems: pUBllO. pCl94. pSl94. 
pSA2100. pEl94. pTl27, pUBll2. pC221. 
pC223. and pABl24 B. subtilis strains RUB 
331 and BCSC 1S53 have been certiFied as the 
host component of HVl systems based on 
these plasmids. 
HV2 — The asporogenic mutant derivative 
of Bacillus subtilis. ASB 298. with the 
following plasmids as' the vector component: 
pUBllO. pCl94. pSl94. pSA2100. pEl94. 
pTT27. pUBll2. pC221. pC223. and pABl24. 
HV2 — The following sterile strains of 
Saccharomyces cerevisiae. all of which have 
the ste-VC9 mutation. SHYl. SHY2. SHY3. 
and SHY4. The following plasmids are 
certified for use: YIpl. YF.p2. YEp4. YIpS. 
YEp8. YRp7. YEp20. YEp21. YEp24. Ylp2S. 
Ylp28 Ylp27. Ylp28. Ylp29 Ylp30. Ylp31. 
YIP32 and Ylp33. 
EK2 Plasmid Systems. The £ call K-12 
strain chi-177e. The following plasmids are 
certified for use: pSClOl. pMB9. pBR313. 
pBR322. pDM24. pBR327. pCLlOI. pHBl The 
following £. coli/S. cerevisiae hybrid 
plasmids are certified as F.K2 vectors when 
used in £ coli chi-1776 or in the sterile yeast 
strains. SMYl SMY2. SHY3. and SHY4 YIpl. 
YF.p2. YEp4 YlpS. YEp6. YRp7. YEp20 YEp21 
YF.p24 Ylp25. Ylp28. Ylp27. Ylp28. Ylp29 
Ylp30. Ylp3t. Ylp3i Ylp33 
EK2 Bdcteriophdge Systems The 
following are certified FK2 systems 
ba.sed on bacteriophage lambda: 
Vrrior 
Xxitve<; xb^ 
X|KU£V XB 
XSiZ|r<r XB 
X<tAlX) AB 
Charon 3 A 
Charrm 4 \ 
Himi 
OPSOSK.if 
DCjOauph 
£- I.’h K-W 
tllTiOsi/nK 
Oeaior lll*3<V.() + 
tH'sn m 
Charon 16A 
Charon ZIA 
Charon 23A 
Charon 24A 
DP50 or DPSOsupF 
DPSOsupF 
DPSO or DPSOsupF 
DP50 or DPSOsupF 
Appendix G.— Containment Conditions for 
Cloning of Genes Coding for the Biosynthesis 
of Toxins for Vertebrates 
1. General Information. Appendix G 
specifies the containment to be used for the 
deliberate cloning of genes coding for the 
biosynthesis of toxins for vertebrates. 
Cloning of genes coding for toxins for 
vertebrates that have an LOm of less than 100 
nanograms per kilogram body weight (e g., 
the botulinum toxins, tetanus toxin, 
diphtheria toxin. Shigella dysenteriae 
neurotoxin) is prohibited. No specific 
restrictions shall apply to the cloning of genes 
if the protein specified by the gene has an 
LOm of 100 micrograms or more per kilogram 
of body weight. Experiments involving genes 
coding for toxins with an LDm of 100 
micrograms or less per kilogram body weight 
shall be registered with ORDA prior to 
initiating the experiments. A list of toxins 
classified as to LOm is available from ORDA. 
Testing procedures for determining toxicity of 
toxins not on the list are available from 
ORDA. The results of such tests shall be 
forwarded to ORDA. which will consult with 
the ad hoc Working Croup on toxins prior to 
inclusion of the toxin on the list. (See Section 
IV-E-l-b-(3)-(i).) 
2. Containment Conditions for Cloning of 
Toxin Cenes in £ coli K-12. 
(a) Cloning of genes coding for toxins for 
vertebrates that have an LD.« in the range of 
100 nanograms to 1000 nanograms per 
kilogram body weight (e g., abrin. Clostridium 
perfringens epsilon toxin) may proceed under 
P2 4 EK2 or P3 EKl containment conditions. 
(b) Cloning of genes for the biosynthesis of 
toxins for vertebrates with an LDm in the 
range of 1 microgram to 100 micrograms per 
kilogram body weight may proceed under 
Pi 4- EKl containment conditions (e.g.. 
Staphylococcus aureus alpha toxin. 
Staphylococcus aureus beta toxin, ricin. 
Pseudomonas aeruginosa exoioxin A. 
Bordatella pertussis toxin, the lethal factor of 
Bacillus anthracis. the Pasteurella pestis 
murine toxins, the oxygen-labile hemolysins 
such us streptolysin O. and certain 
neurotoxins present in snake venoms and 
other venoms). 
(r) Some enterotoxins are substantially 
more toxic vshi-n administered enterally then 
parentt-rallv. the following enterotoxins shall 
l<e xubjei I to PI ► EKl containment 
conditions: cholera toxin, the heat labile 
toxins of £. coli, Klebsiella, and other related 
proteins that may be identified by 
neutralization with an antiserum mono 
specific for cholera toxin, and the heat stable 
toxins of £. coU and of Yersinia 
enterocolitica. 
3. Containment Conditions for Cloning of 
Toxins Cenes in Organisms Other than £ 
coli K-12. 
Requests involving the cloning of genes 
coding for toxins for vertebrates in host- 
vector systems other than £. coli K-12 will be 
evaluated by ORDA. which will consult with 
the ad hoc working group on toxins. (See 
Section IV-E-l-b-(3Hi) ) 
Appendix H. — Experiments Covered by 
Section III-O 
No experiments currently fall under 
Section 111-0 of the Guidelines. 
Dated: June 22. 1981 
Donald S. Fredrickson, M.D.. 
Director. National Institutes of Health. 
OMB's "Mandatory Information 
Requirements for Federal Assistance Program 
Announcements" (45 FR 39592) requires a 
statement concerning the official government 
programs contained in the Catalog of Federal 
Domestic Assistance. Normally NIH lists in 
its announcements the number and title of 
affected individual programs for the guidance 
of the public. Because the guidance in this 
notice covers not only virtually every NIH 
program but also essentially every federal 
research program in which D.N’A recombinant 
molecule techniques could be used, it has 
been determined to be not cost effective or in 
the public interest to attempt to list these 
programs. Such a list would likely require 
several additional pages. In addition. NIH 
could not be certain that every federal 
program would be included as many federal 
agencies, as well us private organizations, 
both national and international, have elected 
to follow the NIH Guidelines. In lieu of the 
individual program listing. NIH invites 
readers to direct questions to the information 
address above about whether individual 
programs listed in the Catalog of Federal 
Domestic Assistance are affected. 
NIH programs are not covered by O.MB 
Circular A-95 because they f't the description 
of "programs not considered appropriate" in 
Section B-(b)-(4) and (5) of that Circular 
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