Nucleic Acids Research, 1986, Vol. 14, No. 14 5777-5792
© 1986
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Isolation and characterization of bovine and mouse terminal deoxynucleotidyltransferase cDNAs expressible in mammalian cells
1DNAX Research Inst. Molecular and Cellular Biology 901 California Avenue, Palo Alto, CA 94304, USA 2Dept. Biochem., Inst. Developmental Research Aichi Prefecture Colony, Kasugai 480-03 3Dept. Biochem., Primate Inst., Univ. Kyoto Kanrin, Inuyama 4Pharmaceutical Inst., Keio Univ. School of Medicine Shinjuku-ku, Shinanomachi 35, Tokyo, Japan
*To whom correspondence should be addressed
Received March 10, 1986. Revised June 9, 1986. Accepted June 9, 1986.
We have isolated nearly full-length cDNA clones of terminal deoxynucleotidyltransferase (TdT) from calf thymus and mouse lymphoma cDNA libraries. The libraries were constructed using the pcD vector system which permits the expression of cDNA inserts in mammalian cells. The bovine TdT cDNA clone contains an open reading frame coding for 520 amino acids, Mr 59,678. The mouse TdT cDNA clone contains an open reading frame of 1,587 bp, whose translated cDNA encodes a 60,004 dalton protein. The mouse TdT cDNA clone contains 60 bp in the 3' end region of the coding sequence not found in the bovine TdT cDNA sequence, otherwise, the clones share about 80% homology. A possible nuclear-localization-sequence (Pro-Arg-Lys-Lys-Arg-Pro-Arg) was conserved in the N-terminal region in the mouse and bovine cDNA clones. Bovine and mouse cDNAs transfected into C0S7 monkey fibroblasts directed the synthesis of enzymatically active protein of Mr 60,000 which was detected immunologically using polyclonal rabbit antibody against bovine TdT. Bovine TdT expressed in CoS7 cells by nearly full-length cDNA clone was localized in the nucleus and the translational product of pOK103 lacking the nuclear-localizatin-sequence was localized in the cytoplasm.
+Present address: Research Institute for Disease Mechanism and Control, Nagoya University School of Medicine, Showa-ku, Tsurumaicho 65, Nagoya, Japan
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