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Molecular analyses of sulfur-cycling...

Karr, Elizabeth Anne.

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Molecular analyses of sulfur-cycling prokaryotes and Archaea in permanently frozen Antarctic lakes.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Molecular analyses of sulfur-cycling prokaryotes and Archaea in permanently frozen Antarctic lakes./
作者:	Karr, Elizabeth Anne.
面頁冊數:	147 p.
附註:	Source: Dissertation Abstracts International, Volume: 64-09, Section: B, page: 4188.
Contained By:	Dissertation Abstracts International64-09B.
標題:	Biology, Microbiology. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=3105702

Molecular analyses of sulfur-cycling prokaryotes and Archaea in permanently frozen Antarctic lakes.
Karr, Elizabeth Anne.

Molecular analyses of sulfur-cycling prokaryotes and Archaea in permanently frozen Antarctic lakes. - 147 p.

Source: Dissertation Abstracts International, Volume: 64-09, Section: B, page: 4188.

Thesis (Ph.D.)--Southern Illinois University at Carbondale, 2003.

The permanently-frozen lakes of the Dry Valleys in Antarctica represent one of the most extreme environments on Earth. Although extensive geochemical data exist for these lakes, few studies examining the microbial diversity of this environment have been undertaken. In this work we examined the biodiversity of sulfur-cycling prokaryotes in Lake Fryxell, a permanently-frozen stratified lake located in the Dry Valleys. In addition, a brief investigation of the purple anoxyphototrophs was undertaken on Lake Hoare. Lake water from various depths was filtered to concentrate the biomass and subjected to genomic DNA and RNA isolation. The genomic DNA was analyzed by polymerase chain reaction (PCR) using primer sets targeting key metabolic genes and/or the gene encoding 16S rRNA. Purple anoxygenic phototrophs were targeted using the pufM gene encoding a component of the photoreaction complex, while the sulfate-reducing bacteria were targeted using the dsrA gene that encodes the alpha-subunit of dissimilatory sulfite-reductase. Archaea were targeted using a 16S rDNA primer set specific to that domain. The resulting PCR products were then analyzed using denaturing gradient gel electrophoresis (DGGE) followed by cloning and sequencing of unique phylotypes.Subjects--Topical Terms:

1017734
Biology, Microbiology.

Molecular analyses of sulfur-cycling prokaryotes and Archaea in permanently frozen Antarctic lakes.
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245 1 0 $a Molecular analyses of sulfur-cycling prokaryotes and Archaea in permanently frozen Antarctic lakes.
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502 $a Thesis (Ph.D.)--Southern Illinois University at Carbondale, 2003.
520 $a The permanently-frozen lakes of the Dry Valleys in Antarctica represent one of the most extreme environments on Earth. Although extensive geochemical data exist for these lakes, few studies examining the microbial diversity of this environment have been undertaken. In this work we examined the biodiversity of sulfur-cycling prokaryotes in Lake Fryxell, a permanently-frozen stratified lake located in the Dry Valleys. In addition, a brief investigation of the purple anoxyphototrophs was undertaken on Lake Hoare. Lake water from various depths was filtered to concentrate the biomass and subjected to genomic DNA and RNA isolation. The genomic DNA was analyzed by polymerase chain reaction (PCR) using primer sets targeting key metabolic genes and/or the gene encoding 16S rRNA. Purple anoxygenic phototrophs were targeted using the pufM gene encoding a component of the photoreaction complex, while the sulfate-reducing bacteria were targeted using the dsrA gene that encodes the alpha-subunit of dissimilatory sulfite-reductase. Archaea were targeted using a 16S rDNA primer set specific to that domain. The resulting PCR products were then analyzed using denaturing gradient gel electrophoresis (DGGE) followed by cloning and sequencing of unique phylotypes.
520 $a Prokaryotes potentially contributing to the sulfur-cycle were detected throughout the water column of Lake Fryxell in both oxic and anoxic zones by PCR of the appropriate gene for each group of sulfur-cycling prokaryotes. Subsequent DGGE analysis of the amplification products revealed the presence of multiple phylotypes at each depth. Phylogenetic analysis of representative phylotypes exhibited various levels of sequence similarity to several purple anoxygenic phototrophs, gram-negative and gram-positive sulfate-reducing bacteria, as well as sulfur-respiring Archaea. In addition, metabolic activity and overall viability were established for the purple anoxyphototrophs and Archaea using reverse-transcription of pufM mRNA and Archaeal 16S rRNA.
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