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Cloning and expression of mitochondrial translational elongation factor Ts from bovine and human liver
H. Xin; V. Woriax; W. Burkhart; L.L. Spremulli (Profiled Author: Velinda L. Woriax)
Journal of Biological Chemistry. 1995;270(29):17243-17249.Abstract
The sequences of the cDNAs for the mitochondrial translational elongation factor Ts (EF-Ts(mt)) from bovine and human liver have been obtained. The deduced amino acid sequence of bovine liver EF-Ts(mt) is 338 residues in length and includes a 55-amino acid signal peptide and a mature protein of 283 residues. The sequence of the mature form of bovine EF-Ts(mt) is 91% identical to that of human EF-Ts(mt) and 29% identical to Escherichia coli EF-Ts. Southern analysis indicates that there are two genes for EF-Ts(mt) in bovine liver chromosomal DNA. A 224-base pair intron is located near the 5'- end of at least one of these genes. Northern analysis using a human multiple tissue blot indicates that EF-Ts(mt) is expressed in all tissues, with the highest levels of expression in skeletal muscle, liver, and kidney. Both the mature and precursor forms of bovine liver EF-Ts(mt) have been expressed in E. coli as histidine-tagged proteins. The mature form of EF-Ts(mt) forms a complex with E. coli elongation factor Tu. This complex is active in poly(U)- directed polymerization of phenylalanine. The precursor form is expressed as a 42-kDa protein, which is rapidly degraded in the cell.
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