Cloning and expression of human tyrosine aminotransferase cDNA.
Complementary DNA clones encoding human tyrosine aminotransferase (TAT) were isolated by screening a normal adult woman liver lambda gt11 library with rat TAT cDNA. The largest isolated cDNA is 2051 bp long (EMBL accession number X55675). This cDNA was subcloned downstream of the cytomegalovirus promoter in the pCMV vector for transfection into human cervical carcinoma HeLa cells. Expression of the TAT cDNA resulted in the synthesis of a protein with a molecular mass of approximately 50 kDa, as assessed by Western analysis, a value which is in close agreement with the predicted molecular weight of 50,399, for a deduced sequence of 454 amino acids. The expressed protein catalyzed specifically the conversion of L-[14C]tyrosine into p-[14C]hydroxyphenylpyruvate. The availability of a functional TAT cDNA provides a useful tool for detailed study of the structure-function relationship of the enzyme and its mutated derivatives.
Séralini
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Medical Research Council Group in Molecular Endocrinology, CHUL Research Centre, Quebec, Canada.
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D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015153:Blotting, Western; D003001:Cloning, Molecular; D018076:DNA, Complementary; D005260:Female; D006367:HeLa Cells; D006801:Humans; D007106:Immune Sera; D008297:Male; D008969:Molecular Sequence Data; D051381:Rats; D014444:Tyrosine Transaminase / Q000235:genetics*
D018076:DNA, Complementary; D007106:Immune Sera; D014444:Tyrosine Transaminase