Actions 
The Guidelines require that major actions taken by the Director of NIH on 
topics relating to recombinant DNA research are to be published in this 
Bulletin. In the Issues Pending section, topics which are currently under 
discussion are briefly described. For example, they may include issues 
which are being examined by the Office of Recombinant DNA Activities or 
the NIH Recombinant DNA Advisory Committee. In the Issues Resolved 
section are presented the actions taken by NIH on issues which have been 
pending. These will be presented in their entirety as printed in the Federal 
Register or in memos to the Institutional Biosafety Committees. 
Issues Pending 
The NIH Recombinant DNA Advisory Committee 
at its last meeting was requested to advise the NIH 
on the following issues. The actions taken by NIH 
will be reported in the next issue of the Bulletin. 
1. The cloning of DNAfrom higher eukaryotes into 
EK2 systems and the return of the cloned DNA 
to higher eukaryotes under appropriate physi¬ 
cal containment conditions. 
2. The cloning, under PI physical containment, of 
DNA into invertebrate cells in culture using 
organelle, plasmid or chromosomal DNA as 
vectors. 
3. The cloning, under PI physical containment, of 
DNA into vertebrate cells in culture using orga¬ 
nelle, plasmid or chromosomal DNA as vectors. 
4. Criteria for approval of lower eukaryotes as 
HVI & HV2 systems. Specifically requested for 
HV1 are laboratory strains of Saccharomyces 
cerevisiae and modified strains of Neurospora 
crassa. 
5. A request by Dr. David Botstein, et. al., for 
approval of a yeast HV2 system based on loss 
of mating abililty. 
6. The use of unmodified N. crassa as HVI sys¬ 
tems when higher levels of physical contain¬ 
ment are used. 
7. New lists of organisms which have been shown 
to exchange DNA and which, therefore, should 
be exempt from the Guidelines (when experi¬ 
ments only involve pairs of organisms on these 
lists). One list involves certain species of Bacil¬ 
lus and the other involves certain pairs of Strep- 
tomyces species. 
8. The approval of asporogenic mutant deriva¬ 
tives of B. subtilis as HV1 systems using spe¬ 
cific plasmids. 
9. The use of the P3 level of containment in those 
cases where two bacterial species do not 
exchange DNA. 
10. The levels of containment for the shotgun clon¬ 
ing of primate and other mammalian DNA can 
be P3 + EK1 as an alternative to the present P2 
+ EK2. 
11. The cloning in B. subtilis under P2 conditions of 
DNA from E. coli, S. cerevisiae and Streptomy- 
ces coellcolor. 
12. The cloning in Streptomyces coellcolor under 
P2 conditions of DNA from B. subtilis, E. coli, 
and Staphylococcus aureus plasmids. 
13. The use of Ff single-strand bacteriophages 
(such as Ml 3) as vectors with E. coli K-12 for 
EK1 containment. 
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