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PMID: 6324091 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Molecular cloning, sequence analysis and in vitro expression of a rat tRNA gene cluster.

Nucleic acids research ·Vol. 11 ·No. 24 ·1983-12-20 ·Pages 8609-24

Makowski DR, Haas RA, Dolan KP, Grunberger D

Abstract

A rat genomic DNA fragment containing a tRNA gene cluster was isolated from a lambda phage library. Hybridization and nucleotide sequence analysis revealed the presence of a 83 bp tRNALeuCUG gene and a 72 bp tRNAAspGUG gene. Both genes possessed intact coding regions and putative transcription termination signals at their respective 3' ends. In vitro transcription analysis of the two subcloned genes in a HeLa cell S-100 system demonstrated the specific synthesis of a number of RNAs by RNA polymerase III. Studies carried out in the presence of alpha-amanitin showed that the larger RNAs are precursors for the final processed transcripts of the tRNALeu and tRNAAsp genes, respectively. Further nucleotide sequence analysis of the cluster revealed the presence of tRNAGly and a tRNAGlu pseudogenes with missing areas within their coding regions which are essential for transcription by RNA polymerase III. Within the region of DNA between the tRNALeu and tRNAAsp genes is a sequence which is 65% homologous to a region of the rat B1 element. The significance of this latter structure within the gene cluster is unknown.

MeSH Terms
Animals Bacteriophage lambda/genetics Base Sequence Cloning, Molecular DNA Restriction Enzymes Escherichia coli/genetics Genes HeLa Cells/metabolism Humans Nucleic Acid Conformation Nucleic Acid Hybridization Plasmids RNA, Transfer/genetics Rats Transcription, Genetic
Chemicals
RNA, Transfer DNA Restriction Enzymes
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Makowski D R
Haas R A
Dolan K P
Grunberger D
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36 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/11.24.8609
Published
1983-12-20
Pages
8609-24
Language
English
Region
England
NLM ID
0411011
PMCID
PMC326611
Subset
IM
Grants
NCI NIH HHS · CA31696 · United States
NCI NIH HHS · F32 CA06897 · United States
Databases
GENBANK
X00218, X00219, X00220
External Links
PubMed source DOI: 10.1093/nar/11.24.8609 PMC Full Text View journal details
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