Isolation and sequence analysis of a human $tRNA^ser$ gene

Abstract

A human $tRNA^{ser}$ gene was found in a sequenced human chromosomal DNA fragment, and secondary structure of $tRNA^{ser}$ was deduced from the gene. The human $tRNA^{ser}$ and secondary structure of $tRNA^{ser}$ was deduced from the gene. The human $tRNA^{ser}$ gene has anticodon of 5``-AGA-3``, and sequence of the gene differs from the previously sequenced human $tRNA^{ser}$ gene in only a 5`` position base of anticodon. This gene had been isolated from human genomic library through plaque hybridization by using labeled human $tRNA^{ser}$ as probe. Here, 1.1Kb DNA fragment was isolated from the DNA of the recombinant $\lambda$ charon 4A which was named $\lambda$ StlB. The DNA fragment was ligated with pUC9 DNA vector, and used to transform E. coli JM83 cells. Recombinant pUC9 DNA from transformant was confirmed to contain tRNA coding gene by dot hybridization with labeled human total tRNA. After restriction mapping, the restrictly digested 1.1Kb fragment was subcloned to M13 RF DNA and M13 singlestranded DNA was obtained for sequencing of DNA. The nucleotides of DNA fragments were sequenced by dideoxynucleotide chain termination sequencing method.