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UBC Theses and Dissertations
Characterization of recombinant plasmids carrying Drosphila melanogaster tRNA Serine7 genes and their preparation for DNA sequencing Spurr, Mark Gregory
Abstract
Specific Drosphila tRNA Ser4,7 plasmids were identified and isolated by hybridization with purified [¹²⁵I] tRNA Ser4,7 molecules. Seven clones were isolated carrying the Drosphila Ser tRNA Ser4,7 gene and were further characterized by restriction endonuclease digestion; agarose gel electrophoresis and hybridization with individual purified [¹²⁵I] tRNA Ser4,7 molecules. The results show that five different DNA fragments have been isolated, four which code for a single, specific isoacceptor, and one which appears to code for two different isoacceptors. Two plasmids which initially contained multiple Hind III inserts upon primary isolation were recloned to contain single Hind III inserts containing the tRNA Ser4,7 gene. One of these recloned plasmids contained a smaller tRNA Ser4,7 gene carrying insert than did its original multiple insert isolate. Small tRNA Ser4,7 gene carrying restriction fragments were labelled with T4 polynucleotide kinase and [³²P] ATP, strand separated, and electroeluted, in preparation for nucleotide sequencing.
Item Metadata
Title |
Characterization of recombinant plasmids carrying Drosphila melanogaster tRNA Serine7 genes and their preparation for DNA sequencing
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Creator | |
Publisher |
University of British Columbia
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Date Issued |
1979
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Description |
Specific Drosphila tRNA Ser4,7 plasmids were identified and
isolated by hybridization with purified [¹²⁵I] tRNA Ser4,7 molecules. Seven clones were isolated carrying the Drosphila Ser
tRNA Ser4,7 gene and were further characterized by restriction
endonuclease digestion; agarose gel electrophoresis and hybridization with individual purified [¹²⁵I] tRNA Ser4,7 molecules. The results show that five different DNA fragments have been isolated, four which code for a single, specific isoacceptor, and one which appears to code for two different isoacceptors. Two plasmids which initially contained multiple Hind III inserts upon primary isolation were recloned to contain single
Hind III inserts containing the tRNA Ser4,7 gene. One of these
recloned plasmids contained a smaller tRNA Ser4,7 gene carrying
insert than did its original multiple insert isolate. Small
tRNA Ser4,7 gene carrying restriction fragments were labelled
with T4 polynucleotide kinase and [³²P] ATP, strand separated, and electroeluted, in preparation for nucleotide sequencing.
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Genre | |
Type | |
Language |
eng
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Date Available |
2010-03-22
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Provider |
Vancouver : University of British Columbia Library
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Rights |
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.
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DOI |
10.14288/1.0095019
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URI | |
Degree | |
Program | |
Affiliation | |
Degree Grantor |
University of British Columbia
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Campus | |
Scholarly Level |
Graduate
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Aggregated Source Repository |
DSpace
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Rights
For non-commercial purposes only, such as research, private study and education. Additional conditions apply, see Terms of Use https://open.library.ubc.ca/terms_of_use.