Expression of simian virus 40 small t antigen in Escherichia coli and purification of the antigen.

S. Ikeda, K. Akiyama, Fumihiro Mitsunobu, T. Wada, M. Hatsushika, S. Watanabe, K. Tsutsui, S. Seki, T. Oda

Research output: Contribution to journalArticle

Abstract

A simian virus 40 (SV 40) DNA fragment encoding small t antigen was cloned in expression vector pUC8 for the purification of the antigen. The SV40 Hind III B fragment was inserted into the Hind III site of pUC8. A plasmid having the lacZ' and small t antigen genes in the same orientation was designated as pSVt. pSVt encodes the entire small t antigen and an extra 18 amino acids at the amino terminus of the antigen. E. coli transformed with pSVt produced hybrid small t antigen (22 kDa) which comprised about 6% of the total protein. The hybrid small t antigen reacted in immunoblot analysis with SV40-induced tumor-bearing hamster serum. Hybrid small t antigen was extracted from E. coli, and purified by preparative SDS-PAGE. The antigen was extracted from the gel using formic acid solution with high-yield. The gel-purified antigen showed the same antigenic reactivity as crude antigen.

Original languageEnglish
Pages (from-to)275-281
Number of pages7
JournalArchiv fur Geschwulstforschung
Volume57
Issue number4
Publication statusPublished - 1987

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Simian virus 40
Escherichia coli
Antigens
formic acid
Gels
Cricetinae
Polyacrylamide Gel Electrophoresis
Plasmids

ASJC Scopus subject areas

  • Cancer Research

Cite this

Ikeda, S., Akiyama, K., Mitsunobu, F., Wada, T., Hatsushika, M., Watanabe, S., ... Oda, T. (1987). Expression of simian virus 40 small t antigen in Escherichia coli and purification of the antigen. Archiv fur Geschwulstforschung, 57(4), 275-281.

Expression of simian virus 40 small t antigen in Escherichia coli and purification of the antigen. / Ikeda, S.; Akiyama, K.; Mitsunobu, Fumihiro; Wada, T.; Hatsushika, M.; Watanabe, S.; Tsutsui, K.; Seki, S.; Oda, T.

In: Archiv fur Geschwulstforschung, Vol. 57, No. 4, 1987, p. 275-281.

Research output: Contribution to journalArticle

Ikeda, S, Akiyama, K, Mitsunobu, F, Wada, T, Hatsushika, M, Watanabe, S, Tsutsui, K, Seki, S & Oda, T 1987, 'Expression of simian virus 40 small t antigen in Escherichia coli and purification of the antigen.', Archiv fur Geschwulstforschung, vol. 57, no. 4, pp. 275-281.
Ikeda, S. ; Akiyama, K. ; Mitsunobu, Fumihiro ; Wada, T. ; Hatsushika, M. ; Watanabe, S. ; Tsutsui, K. ; Seki, S. ; Oda, T. / Expression of simian virus 40 small t antigen in Escherichia coli and purification of the antigen. In: Archiv fur Geschwulstforschung. 1987 ; Vol. 57, No. 4. pp. 275-281.
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AU - Wada, T.

AU - Hatsushika, M.

AU - Watanabe, S.

AU - Tsutsui, K.

AU - Seki, S.

AU - Oda, T.

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