• Title/Summary/Keyword: Human Poly(ADP-ribose)synthetase cDNA

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Molecular Cloning and Expression of Human Poly (ADP-ribose) Synthetase cDNA in E. Coli (인간 Poly(ADP-ribose) Synthetase cDNA의 클로닝 및 대장균에서의 발현)

  • 이성용;김완주;이태성;박상대;이정섭;박종군
    • The Korean Journal of Zoology
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    • v.39 no.3
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    • pp.248-256
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    • 1996
  • The present study was performed to clone and express human poly (ADP-ribose) synthetase (PARS) cDNA in E coli. For these purposes, the CDNA for human poly (ADP-ribose) synthetase, encoding the entire protein, was cloned into pGEM-7Zf(+). The resulting recombinant plasmid pPARS6.1 was restriction enzyme mapped and its identity was confirmed by Southern blot analysis. The pPARS6. 1 contained full-length CDNA of human PARS and the nudeotide sequences were identical with those reported previously. The recombinant protein which migrated as a unique 120 kDa band on 10% SDS-polyacrylamide gels, was identified as PARS by Southwestern blots using nick-translated DNA probes and by activity gels and activity blots using 32 P-NAD as a substrate for poly (ADP-ribose) synthetase (PARS). The signals corresponding to 120 and 98 kDa proteins were obtained following IPTG (0.4 mM) induction of the PARS cDNA cloned into Xba I-digested pGEM-7Zf(+) vector. Nonspecific signals corresponding to 45 and 38 kDa proteins were also shown in both IPTG-induced and noninduced cells. The nonspecific proteins may be products of incomplete translation or proteolytic products of intact PARS.

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