M.S. Rudinski, and D.H. Dean, in preparation). 
Fragment band shifts between pi 4 and odo7 are 
shown in Fig. 2. These cloning vehicles provide 
the mechanism for integrating cloned DNA by 
insertion into the host chromosome (to achieve 
low copy allelic complementation), or lytic cycle 
amplification/expression of cloned DNA during 
the lytic cycle. 
4. Demonstration of Cloning. According to pres¬ 
ent indications, (fdo? vehicles carrying cloned 
DNA make a discernably lighterturbid plaquethan 
(t)do7. Both agarose gel restriction analysis and 
heteroduplex analysis of phage from such 
plaques, called x phages, reveal cloned DNA is 
carried. BcoRI analysis of x5 (Fig. 2) shows an 
absence of the F band and appearance of a new 
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1. Ehrlich, S.D,, DNA cloning In Bacillus subtilis, Proc. Nat. 
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of chimeric plasmids in Bacillus subtilis. Proc. Nat. Acad. Sci 
USA 75: 1428-1432, 1978. 
3. Keggins, K.M., Lovett, P S., and Duvall, E.J., Molecular clon¬ 
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4. Kraft, J., Bernhard, K., and Goebel, W., Recombinant plas¬ 
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8, Murray, K. and Murray. N.E., Manipulation of restriction 
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9. Thomas M., Cameron. J.R., and Davis, R.W,, Viable molecu¬ 
higher molecular weight band. SincetheSa/GI site 
is within the EcoRI F band, this result is consistent 
with extra DNA inserted into the Sa/GI site. Fig. 3 
(data of J.B. Perkins, J.M. Kroyer, and D.H. Dean, 
manuscript in preparation) shows heteroduplex 
molecules of x5 and pi 4. Arrows indicate the 
inserted DNA (R-DNA) and the (j)do7 deletion 
loop(A). X 5 carries 0.5 x 10^ dalton of identified 
cloned DNA, Current work involves removal of all 
but one EcoRI site (the one joining fragment A and 
F) which would allow cloning with EcoRI and the 
insertion of an adaptor molecule for SamHI into 
the Sa/GI or EcoRI site. 
This phage cloning vehicle in conjunction with 
the currently available plasmid cloning vehicles 
(1,2,3,4,5) will provide Recombinant DNA systems 
for further genetic studies of B. subtilis. 
lar hybrids of bacteriophage lambda and eucaryotic DNA, 
Proc, Natl, Acad, Sci. USA 71 4579-4583, 1974, 
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temperate phages of Bacillus subtilis. Cur. Micro., 1: 213-217, 
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13. Dean, D.H,, Orrego, J.C,, Hutchison, K,W,, and Halvorson, 
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of Virol. 20: 509-519, 1976, 
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endonuclease mapping of bacteriophage |l05 and closely 
related temperate Bacillus subtilis bacteriophages plO and 
pi 4, J. of Virol. 28: 403-407, 1978. 
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perate bacteriophage molecular vehicle for Bacillus subtilis. 
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16. Ainsworth, G,C, and Sneath, P.H A., Microbial classifica¬ 
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