Isolation of tRNA from uropathogenic escherichia coli

ABSTRACT

Transfer RNA is a type of RNA which during protein synthesis Act as an adaptor molecule, matching amino acids to their codons on mRNA. tRNA also functions in the formation of cross links during peptidoglycan synthesis. The aim of this study is that, extraction of tRNA from uropathogenic Escherichia coli then detect the presence of such molecules after extraction and measure the purity of the tRNA extract solutions. Thirty uropathogenic E.coli isolates were isolated from hospitalized and non hospitalized patients, complaining of urinary tract infections, of Al-Kadhymia Teaching Hospital and subjected to tRNA extraction. A method of tRNA extraction was modified by adding sodium dodecyl sulfate [SDS] instead of urea. Polyacrylamide gel electrophoresis and two methods of staining, ethidium bromide staining and silver staining, as well as spectrophotometric detection were used. Ethidium bromide stained gel reveals bands with molecular weight less than yeast rRNA .Silver stained gel shows bands with molecular weight less than ova albumin[45000 dalton] but with approximate molecular weight of chymotrpsinogen[24000 dalton]. The tRNA extracts were relatively pure with the modified method of extraction. In the present study, a modification of polyacrylamide gel electrophoresis to detect tRNA and to determine their molecular weigh was applied