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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
4
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pubmed:dateCreated |
1988-3-21
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pubmed:abstractText |
A recombinant plasmid was constructed with six synthetic DNA oligomers such that the DNA sequence corresponding to yeast tRNA(Phe) is flanked by a T7 promoter and a BstNI restriction site. Runoff transcription of the BstNI-digested plasmid with T7 RNA polymerase gives an unmodified tRNA of the expected sequence having correct 5' and 3' termini. This tRNA(Phe) transcript can be specifically aminoacylated by yeast phenylalanyl-tRNA synthetase and has a Km only 4-fold higher than that of the native yeast tRNA(Phe). The Km is independent of Mg2+ concentration, whereas the Vmax is very dependent on Mg2+ concentration. Comparison of the melting profiles of the native and the unmodified tRNA(Phe) at different Mg2+ concentrations suggests that the unmodified tRNA(Phe) has a less stable tertiary structure. Using one additional DNA oligomer, a mutant plasmid was constructed having a guanosine to thymidine change at position 20 in the tRNA gene. A decrease in Vmax/Km by a factor of 14 for aminoacylation of the mutant tRNA(Phe) transcript is observed.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-1264199,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-247991,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-333395,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-343112,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-3550694,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-357743,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-36738,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-3684574,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-377228,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-3846456,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-3894009,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-4080546,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-4553754,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-4586558,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-6371001,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-6371808,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-6472477,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-6553357,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-6728685,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-692735,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-7003543,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-7004643,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-7016875,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-7029537,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-7041969,
http://linkedlifedata.com/resource/pubmed/commentcorrection/3277187-790568
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Fungal,
http://linkedlifedata.com/resource/pubmed/chemical/DNA, Recombinant,
http://linkedlifedata.com/resource/pubmed/chemical/Phenylalanine,
http://linkedlifedata.com/resource/pubmed/chemical/Phenylalanine-tRNA Ligase,
http://linkedlifedata.com/resource/pubmed/chemical/RNA, Fungal,
http://linkedlifedata.com/resource/pubmed/chemical/RNA, Transfer, Amino Acid-Specific,
http://linkedlifedata.com/resource/pubmed/chemical/RNA, Transfer, Phe
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pubmed:status |
MEDLINE
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pubmed:month |
Feb
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pubmed:issn |
0027-8424
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
85
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
1033-7
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:3277187-Base Sequence,
pubmed-meshheading:3277187-DNA, Fungal,
pubmed-meshheading:3277187-DNA, Recombinant,
pubmed-meshheading:3277187-Molecular Sequence Data,
pubmed-meshheading:3277187-Nucleic Acid Conformation,
pubmed-meshheading:3277187-Nucleic Acid Denaturation,
pubmed-meshheading:3277187-Phenylalanine,
pubmed-meshheading:3277187-Phenylalanine-tRNA Ligase,
pubmed-meshheading:3277187-RNA, Fungal,
pubmed-meshheading:3277187-RNA, Transfer, Amino Acid-Specific,
pubmed-meshheading:3277187-RNA, Transfer, Phe,
pubmed-meshheading:3277187-Saccharomyces cerevisiae
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pubmed:year |
1988
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pubmed:articleTitle |
Biochemical and physical characterization of an unmodified yeast phenylalanine transfer RNA transcribed in vitro.
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pubmed:affiliation |
Department of Chemistry and Biochemistry, University of Colorado, Boulder 80309.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.
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