Primary structure of alanyl-tRNA synthetase and the regulation of its mRNA levels in Bombyx mori.

cDNA clones encoding Bombyx mori alanyl-tRNA synthetase were isolated from a library in lambda gt11 using antibody, synthetic oligonucleotides, and a characterized cDNA as probes. Analysis of the sequence revealed significant homology between the B. mori and Escherichia coli alanyl-tRNA synthetases, particularly in their amino-terminal domains. Northern blot analysis indicated that the mRNA for alanyl-tRNA synthetase is 3.8 kilobase pairs in mRNA isolated from posterior silk gland, middle silk gland, and ovarian tissue. Steady-state levels of alanyl-tRNA synthetase mRNA in the posterior silk gland increased in the first 48 h of the fifth larval instar, decreasing gradually thereafter. In the middle silk gland, alanyl-tRNA synthetase mRNA peaked at 72 h of the fifth larval instar, declining to undetectable levels by 120 h. Genomic Southern blot analysis using a nick-translated cDNA probe revealed hybridization to single fragments when B. mori genomic DNA was digested with various restriction endonucleases.