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Related post: Project No. ZOl AI 00006-09 LSD
established plasmids are amplifiable with chloramphenicol.
Because of the interest in streptococci as cloning vehicles, I
plan to continue this work and hope to elucidate the plasmid
Spiroplasma plasmids . Prior work with W. O. Mitchell, T. J.
Popkin, and R. M. Cole (See Flutamide Tablets Projects ZOl AI 00007-06 LSD and ZOl
AI 00004-20 LSD) was extended, confirmed, and published. Present
data show that 12 Flutamide Hair Loss of 14 spiroplasmas carry from 1 to 8 cryptic
plasmids per strain. Sizing by both electron microscopy and
agarose gel electrophoresis (the latter using new standards) g
showed a range of molecular weights between 1 x 10 and 26 x 10
daltons. Several size classes occurred in common in different
spiroplasma strains. The amount of plasmid DNA per strain was as
much as 12% of total cellular DNA, as determined by radioactive
labelling. No such multiplicity of plasmids, with some in common
among several strains, were previously known in any mycoplasmas,
and their newly-recognized presence in spiroplasmas raises
questions of their relation to phenotypic traits, that have yet to
be answered. Preliminary studies showed that a non-helical "mutant"
with a distinctive colonial morphology lacks several plasmids
present in the parent strain, but trials of plasmid curing of the
letter by use of intercalating agents to produce "mutant" colonies
have not yet been successful. Future work will be directed to
this and other approaches to determining the functions of
spiroplasma plasmids, and to their detection and characterization
in strains not yet examined.
Streptococcal bacteriophages . Because of the paucity of current
information. Dr. Moynet (in collaboration with A. colon-Whitt: ZOl
AI 00005-15; See also ZOl AI 00004-20) initiated studies to
characterize several phages. Nine phages, of which 3 are virulent
and 6 temperate, from streptococci of serologic groups A, C, G, E,
and H, are being compared. Seven are long-tailed polyhedral
phages (Type B) as are most streptococcal phages, but 2 are short-
tailed with accessory fibers. DNAs from all have been prepared,
and are being analyzed directly for size, Flutamide 250 Mg after denaturation and
renaturation for the presence of circular permutations, and after
exonuclease III treatment for the presence of terminal repetitions
of nucleotide sequences. Early results suggest smaller molecular
size and absence of permutation in DNA of short- tailed phages.
Other studies in progress include comparison of restriction enzyme
cleavage patterns to determine regions of homology in the DNAs,
and gel electrophoretic comparison of capsid proteins.
Dr. Moynet has also continued physical mapping of the genome (a
24.1 Mdal linear ds-DNA) of Group H streptococcal phage 042.
Project No. ZOl AI 00006-0 9 LSD
Because of the previously shown presence of circular permutation,
the map — when completed — will be circular. By the use of single
and double digests with restriction endonucleases (chiefly Sal I
and Pvu II) , followed by ordering and sizing of resultant
fragments by agarose gel electrophoresis, and of ^treatments with
large fragment Polymerase I in the presence of [ P] -thymidine
triphosphate, it was shown that ^42 DNA possesses 2 Sal I sites
that encompass a Pvu II site. One of these now- identified Sal I
sites has been chosen arbitrarily as the origin, and construction
of the map will proceed.
Hemophilus bacteriophage Physical mapping of the genome of 0N3
of H. influenzae was partially completed in 1977-78 (E. Jablonska,
Visiting Fellow: See Project No. ZOl AI 0006-7 LSD) , but ambiguities
existed among small fragments, in the middle of the genome, generated
by restriction endonucleases EcoRl, Xhol, and Xbal. Resolution of
this problem is being approached by Dr. Moynet and Dr. Jablonska
(as a short-term Guest Worker) by filter hybridization of EcoRl
and Xhol fragments with Xbal fragments, and is expected to lead
shortly to completion of the map.
Ranhand, J. M. : DNA binding and uptake by competent Streptococcus
sanguis Wicky cells. In Glover, S., and Butler, L. O. (eds.).
Proc. of the 4th European Meeting on Transformation and Trans-
fection. Cotswold Press, Oxford, 1979, pp. 60-72.
Ranhand, J. M. : Interactions of competent Streptococcus sanguis
(Wicky) cells with native or denatured, homologous or heterologous
deoxyribonucleic acids. J. Bacteriol. 142: 568-580, 1980.
Ranhand, J. M. , Mitchell, W. O. , Popkin, T. J., and Cole, R, M. :
Covalently-closed circular Deoxyribonucleic acids in spiroplasmas .
J. Bacteriol. (In press: Vol. 143, Sept. 1980)
SMITHSONIAN SCIENCE INFORMATION EXCHANGE
PROJECT NUMBER (Oo NOT use this space}
October 1, 197 9 to September 3 0,198
TITLE OF PROJECT (80 characters or less)
Viruses of Spiroplasmas and Mycoplasmas
U.S. DEPARTMENT OF
HEALTH, EDUCATION, AND WELFARE
PUBLIC HEALTH SERVICE
NOTICE Buy Flutamide OF
INTRAMURAL RESEARCH PROJECT
ZOl AI 00007-05 LSD
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