Physical Mapping of the Synechococcus Sp. Strain PCC 7942 Chromosome
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Authors
Ransom, Wendy D.
Issue Date
1991
Type
Thesis
Language
en_US
Keywords
Alternative Title
Abstract
A partial physical map of the Synechococcus sp. strain PCC 7942
chromosome was constructed by the ordering of restriction fragments
resolved with pulse field gel electrophoresis, cosmid-based walking using a
preexisting cosmid library, and gene specific probings of pulse field gels.
Cosmid-based walking on the right side of the rbcL locus was extended 60
kb by probing the library for related cosmids and by examining the
restriction patterns of related cosmids. Problems of pseudo-linking
cosmids to the existing map of the rbcL locus, and identification of cosmids
in two different areas of the chromosome ended mapping at that locus.
Chromosomal DNA was cleaved with Sfi I restriction endonuclease into 11
separable fragments by pulsed field gel electrophoresis. Five of the 11
fragments were linked together through cross hybridization with a gene-specific
probe containing psbAIII, and linking cosmids from the library
were identified on the fragments. The cosmids identified from
hybridization of the fragments with the library revealed groups of cosmids
shared between the fragments. These data indicated that the five
fragments resulted from partial digestion with the enzyme, such that some
of the fragments were overlapping subfragments of the others. A
Synechococcus wox a gene-specific probe was hybridized with the cosmid
library identifying cosmids containing the gene in the chromosome. One of
the cosmids identified was mapped using the restriction endonucleases
MluI, HindIII, BamHI, XbaI, and NotI to determine the position of the
gene on the cosmid. The cosmids containing woxA were recognized by
hybridization of the Sfi 9 fragment with the library and woxA was
positioned within the Sfi 9 fragment through examining restriction
patterns generated from Hind III / EcoRI double digest.
Description
vi, 37 p.
Citation
Publisher
Kalamazoo College
License
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