finitimus, which can be distinguished from other 
varieties of B. thuringiensis by colony and crystal 
spore morphology, were found to produce fS- 
exotoxin and parasporal crystals toxic to the cab¬ 
bage looper. 
This laboratory is currently studying the trans¬ 
formation system in B. thuringiensis var kurstaki 
and B. popilliae using the pUB 110 from Staphyio- 
coccus aureus that mediates kanamycin/neo- 
mycin resistance (Lacey and Chopra, 1974). 
In considering selected entomcpathogenic 
bacteria as hosts for HV-1 systems it is well to 
keep in mindthat underthe new revised guidelines 
new HVTs need not offer a distinct advantage 
over E. coii K-12 host-vectors, need not be capa¬ 
ble of modification to HV2 and HV3, and need not 
be class 1 etiologic agents. B. thuringiensis and B. 
popiliiae enjoy ERA registration and are known to 
be harmless to humans, domestic animals, wild¬ 
life, plants, honeybees, and other non-target orga¬ 
nisms. Neither B. thuringiensis nor B. popilliae 
established themselves in the normal bowel or 
multiply in the alimentary tract, although spores 
remain viable during passage through the intesti¬ 
nal traet. Although 8. popilliae and 8. thuringiensis 
survive only in the spore stage in the environment, 
they only affect specific target pest insects and 8. 
thuringiensis normally does not multiply in the 
target insect. Neither are capable of spreading 
from animal to animal or plant to plant except 
target insects. They do not multiply on body surfa¬ 
ces or intestines and lungs nor penetrate animal 
cells or spread through animal bodies except 
target pest insects. They only produce toxins that 
affect specific pest insects (with the exception of 
the few varieties of 8. thuringiensis producing the 
^-exotoxin which are not approved for use by 
ERA), but otherwise do not alter other living things 
to cause disease. They do not resist normal body 
defense mechanisms, or establish themselves as 
permanent residents of human beings or other 
non-target organisms. 
It would, therefore, appear that these two orga¬ 
nisms meet the requirements for use as host in 
HV-1 systems. 1 propose that the S/aphy/ococcus 
aureus plasmids, which have already been 
approved for cloning DNA in Bacillus subtilis, 
could be used in 8. thuringiensis and 6 . popilliae 
under the same containment conditions. Shotgun 
experiments with DNA's from either 8. thuringien¬ 
sis or 8. popilliae using plasmid vectors from Sta- 
thylococcus aureus or plasmid vectors from, for 
example, 8. thuringiensis into 8. popilliae, or vice 
versa, would be performed under R2 containment. 
Likewise, DNA’s from other specific insect patho¬ 
gens meeting the safety criteria for registration, 
such as Bacillus sphaericus, 8. moritai, or recom¬ 
binant DNA entomopathogenic bacterial phages 
would be performed under R2 physical contain¬ 
ment. Other recombinant DNA experiments using 
8. thuringiensis. 8. popilliae and/or entomopatho¬ 
genic bacteria as hosts that meet the safety crite¬ 
ria for registration would be performed under the 
physical containment requirements specified in 
the guidelines for HV-1 systems. Varieties of 6. 
thuringiensis producing the /3-exotoxin would not 
be allowed for use under these containment 
conditions. 
It is suggestedthat increased recombinant DNA 
research emphasis be focused on these bacteria 
which have some safety advantages and possible 
agricultural benefit over other bacteria currently 
used as recombinant DNA tools and that the ento¬ 
mopathogenic bacteria deemed safe by the Envir¬ 
onmental Rrotection Agency as biological 
insecticides be considered for approval as hosts 
for HV-1 systems. Lastly, it is unlikely that agricul¬ 
ture will best be served through the use of E. coli K- 
12 or 8. subtilis: alternate host-vector systems are 
essential if the potential of recombinant DNA tech¬ 
nology for agriculture is to be realized. 
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