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Federal Register / Vol. 46, No. 210 / Friday, October 30, 1981 / Notices 
"3. Experiments Involving 
Saccharomyces cerevisiae host-vector 
systems. Experiments which use 
Saccharomyces cerevisiae host-vector 
systems, with the exception of 
experiments listed below, are exempt 
from these Guidelines provided that 
laboratory strains .are used. 
“For these exempt experiments. Pi 
physical containment conditions are 
recommended. 
"Exceptions. 
"Experiments described in Section I- 
D-1 to I-D-5 as being prohibited. 
“Experiments involving CDC Class 3 
organisms [1] or cells known to be 
infected with these agents, or any 
recombinant DNA molecules which 
increase the virulence and host-range of 
a plant pathogen beyond that which 
occurs by natural genetic exchange. (See 
Section III-O-l.) 
“Large-scale experiments (e.g., more 
than 10 liters of culture) require prior 
IBC review and approval. 
“Experiments involving the deliberate 
cloning of genes coding for the 
biosynthesis of toxins potent for 
vertebrates. (See Appendix G.) 
“4. Experiments Involving Bacillus 
subtilis host-vector systems. Any 
asporogenic Bacillus subtilis strain 
which does not revert to a sporeformer 
with a frequency greater than 10"’' can 
by used for cloning DNA from any 
nonprohibited source, with the 
exception of those experiments listed 
below. Indigenous Bacillus plasmids 
and phages, whose host-range does not 
include Bacillus cereus or Bacillus 
anthracis, may be used as vectors. 
“For these exempt experiments, PI 
physical containment conditions are 
recommended. 
"Exceptions. 
“Experiments described in Sections I- 
D- to I-D-5 as being prohibited. 
“Experiments involving CDC Class 3 
organisms [1] or cells known to be 
infected with these agents, or any 
recombinant DNA molecules which 
increase the virulence and host-range of 
a plant pathogen beyond that which 
occurs by natural genetic exchange. (See 
Section III-O-l.) 
“Large-scale experiments (e.g., more 
than 10 liters of culture) require prior 
IBC review and approval. 
“Experiments involving the deliberate 
cloning of genes coding for the 
biosynthesis of toxins potent for 
vertebrates. (See Appendix G.)" 
D. Cloning ofSubgenomic Segments of 
Rift Valley Fever Virus 
A new item, munber 36, is added to 
Appendix E: 
“36. Permission is granted to clone in 
E. coli K-12, under Pi physical 
containment conditions, subgenomic 
segments of Rift Valley Fever Virus 
subject to conditions which have been 
set forth by the RAC.” 
E. Addition of Streptococcus Pyogenes 
to Sublist F, Appendix A 
Sublist F, Appendix A, is amended to 
read as follows: 
"Sublist F. 
“1. Streptococcus sanguis. 
“2. Streptococcus pneumoniae. 
“3. Streptococcus faecalis. 
“4. Streptococcus pyogenes. " 
F. Addition of Streptococcus Lactis to 
Sublist E, Appendix A, and Permission 
To Transfer a Recombinant Plasmid 
From S. Faecalisito S. Lactis 
Sublist E, Appendix, is amended to 
read as follows: 
"Sublist E. 
“One way transfer of Streptococcus 
mutans or Streptococcus lactis DNA 
into Streptococcus sanguis . " 
A new entry, item 37, is added to 
Appendix E: 
“37. Permission is given to transfer a 
recombinant lactose plasmid from 
Streptococcus faecalis to S. lactis by 
conjugation.” 
G. Cloning of Saccharomyces 
Cerevisiae DNA in Salmonella 
Typhimurium 
A new entry, number 38, is added to 
Appendix E: 
“38. Attenuated laboratory strains of 
Salmonella typhimurium may be used 
under PI physical containment 
conditions to screen for the 
Saccharomyces cerevisiae 
pseudouridine synthetase gene. The 
plasmid YEpl3 will be employed as the 
vector.” 
H. Cloning Moloney MuLV Provirus and 
Flanking Regions in Hemophilus 
Parainfluenzae 
A new entry, item 39, is added to 
Appendix E: 
“39. Permission is granted to clone in 
Haemophilus parainfluenzae Moloney 
murine leukemia provirus and mouse 
cellular flanking sequences employing 
the plasmid vector, pRK290, under P2 
containment conditions.” 
I. Development of a Host-Vector System 
Based on Corynebacterium Glatamicum 
A new entry, item 40, is added to 
Appendix E: 
“40. Permission is granted for the 
development, under Pi conditions, of a 
new host-vector system based on the 
use of Corynebacterium glutamicum as 
host and non-conjugative poorly 
mobilizable plasmid as vectors.” 
J. Use of Con/ugative Plasmids to 
Transfer DNA Between E. Coli, Vibrio 
Cholera, and Vibrio Harveyi 
A new entry, number 41, is added to 
Appendix E: 
”41. Vibrio harveyi DNA may be 
cloned in Vibrio cholera; plasmids may 
be used to transfer the cloned V. 
harveyi DNA between E. coli K-12, V, 
cholera, and V. harveyi. P2 physical 
containment conditions are required for 
those experiments involving V. cholera. 
Pi containment conditions may be used 
for other phases of the project.” 
K. Containment Conditions for Cloning 
and Expression of DNA Coding for 
Diphtheria Toxin 
Number 30 of Appendix E is modified 
to read as follows: 
“30. Permission is gremted to clone in ' 
E. coli K-12, in high containment 
Building 550 at the Frederick Cancer 
Research Center, restriction fragments 
of Corynephage Beta carrying the 
structural gene for diphtheria toxin. 
Laboratory practices and containment 
equipment are to be specified by the 
IBC.” 
•n 
Additional Announcements of the 
Director, NIAID 
Section IV-E-l-b-(3)-(d) of the 
Guidelines gives responsibility to the 
Director, NIH or his delegate, for 
“authorizing, under procedures specified 
by the RAC, large-scale experiments 
(i.e^ involving more than 10 liters of 
culture) for recombinant DNAs that are 
rigorously characterized and free of 
harmful sequences.” 
Accordin^y, several requests for 
authorization to culture, on a large- 
scale, recombinant DNA host-vector 
systems have been received and 
reviewed by the NIH. 
I. Genentech, Inc. 
On October 21, 1981 the Director, 
NIAID, on the recommendation of the 
RAC, approved requests from 
Genentech, Inc., for the large-scale 
culture of: 
1. E. coli K-12 containing plasmids 
coding for a human calcitonin analog, 
2. Saccharomyces cerevisiae 
containing plasmids into which have 
been ligated cDNA coding for human 
leukocyte interferons A and D, 
3. E. coli K-12 containing plasmids 
coding for porcine growth hormone, 
4. E. coli K-12 containing a plasmid 
into which had been ligated chemically 
synthesized DNA and cloned cDNA 
coding for the VPi protein of Foot and 
Mouth Disease Virus. 
The requests were approved with the 
understanding that Genentech, Inc., has 
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