Abstract

The use of insertion sequence probes for the analysis of fusants obtained following protoplast fusion is described. Hybridization of both total and plasmid DNA from parent and fusant strains with probes to IS904 and ISS1 showed that of the four protoplast fusions examined, three appeared to involve a rearrangement of genetic material while in the fourth the fusant appeared similar to one of the parental strains. This method of analysis provides more information about the changes induced by protoplast fusion than that obtained by monitoring the acquisition or loss of individual characteristics.

References

1
Gasson
M.J.
(
1980
)
Production, regeneration and fusion of protoplasts in lactic streptococci
FEMS Microbiol. Lett.
 ,
9
,
99
102
.
2
Kondo
J.K.
McKay
L.L.
(
1982
)
Transformation of Streptococcus lactis protoplasts by plasmid DNA
Appl. Environ. Microbiol.
 ,
43
,
1213
1215
.
3
Yu
R.S.-T.
Kyle
W.S.A.
Azad
A.A.
Hung
T.V.
(
1984
)
Conditions for the production and regeneration of protoplasts from lactic streptococci
Milchwissenschaft
 ,
39
,
136
139
.
4
Okamoto
T.
Fujita
Y.
Irie
R.
(
1983
)
Protoplast formation and regeneration of Streptococcus lactis cells
Agric. Biol. Chem.
 ,
47
,
259
263
.
5
Simon
D.
Rouault
A.
Chopin
M.-C.
(
1985
)
Protoplast transformation of group N streptococci with cryptic plasmids
FEMS Microbiol. Lett.
 ,
26
,
239
241
.
6
Polzin
K.M.
Shimizu-Kadota
M.
(
1987
)
Identification of a new insertion element, similar to Gram-negative IS26, on the lactose plasmid of Streptococcus lactis ML3
J. Bacteriol.
 ,
169
,
5481
5488
.
7
Schafer
A.
Jahns
A.
Geis
Teuber
M.
(
1991
)
Distribution of the IS elements ISS1 and IS904 in lactococci
FEMS Microbiol. Lett.
 ,
80
,
311
318
.
8
Polzin
K.M.
McKay
L.L.
(
1991
)
Identification, DNA sequence and distribution of IS981, a new, high copy number insertion sequence in lactococci
Appl. Environ. Microbiol.
 ,
57
,
734
743
.
9
Polzin
K.M.
Romero
D.
Shimizu-Kadota
M.
Klaenhammer
T.R.
McKay
L.L.
(
1993
)
Copy number and location of insertion sequences ISS1 and IS981 in lactococci and several other lactic acid bacteria
J. Dairy Sci.
 ,
76
,
1243
1252
.
10
Dodd
M.
Horn
N.
Gasson
M.J.
(
1990
)
Analysis of the genetic determinant for production of the peptide antibiotic nisin
J. Gen. Microbiol.
 ,
136
,
555
566
.
11
Terzaghi
B.E.
Sandine
W.E.
(
1975
)
Improved medium for lactic streptococci and their bacteriophages
Appl. Microbiol.
 ,
29
,
809
813
.
12
Anderson
D.G.
McKay
L.L.
(
1983
)
Simple and rapid method for isolating large plasmid DNA from lactic streptococci
Appl. Environ. Microbiol.
 ,
46
,
549
552
.
13
Southern
E.M.
(
1975
)
Detection of specific sequences among DNA fragments separated by gel electrophoresis
J. Mol. Biol.
 ,
98
,
503
517
.
14
Romero
D.A.
Klaenhammer
T.R.
(
1993
)
Transposable elements in lactococci: a review
J. Dairy Sci.
 ,
76
,
1
19
.