Isolation of yeast tRNALeu genes. DNA sequence of a cloned tRNALeu3 gene.
A library of cloned yeast DNA fragments generated by digestion of yeast DNA with the restriction endonuclease Bam HI has been screened by colony hybridization to total yeast [32P]tRNA. Four hundred colonies carrying yeast tRNA genes were isolated. By hybridization to 125I-tRNALeu3, we have isolated from this collection 14 colonies carrying fragments containing yeast tRNALeu genes. The size of the yeast Bam HI inserts ranged from 2.45 x 10(6) to 14 x 10(6) daltons. One of these fragments was mapped in detail by restriction endonuclease digestion and hybridization to 125I-tRNALeu3. The presence of a tRNALeu3 gene was confirmed by DNA sequence. The results indicate that the tRNALeu3 coding region is not co-linear with the tRNALeu3. An intervening tract of 33 base pairs interrupts the coding sequences 1 base pair past the anticodon coding region. The putative structure of a tRNALeu3 precursor is deduced in which the anticodon base pairs with residues from the intervening sequence.
D001483:Base Sequence; D003001:Cloning, Molecular; D004262:DNA Restriction Enzymes; D004271:DNA, Fungal / Q000302:isolation & purification / Q000378:metabolism; D004274:DNA, Recombinant / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism*; D007930:Leucine; D009690:Nucleic Acid Conformation; D009693:Nucleic Acid Hybridization; D012343:RNA, Transfer / Q000096:biosynthesis*; D012441:Saccharomyces cerevisiae / Q000378:metabolism*
D004271:DNA, Fungal; D004274:DNA, Recombinant; D012343:RNA, Transfer; D004262:DNA Restriction Enzymes; D007930:Leucine