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Nucleic Acids Research, 1991, Vol. 19, No. 7 1571-1576
© 1991

MOLECULAR BIOLOGY

Upstream sequences of rice proliferating cell nuclear antigen (PCNA) gene mediate expression of PCNA-GUS chimeric gene in meristems of transgenic tobacco plants

Shunichi Kosugi, Iwao Suzuka1, Yuko Ohashi2,*, Taka Murakami2 and Yuji Arai

Institute of Applied Biochemistry, University of Tsukuba Tsukuba Science City, Ibaraki 305, Japan 1National Institute of Animal Health Tsukuba Science City, Ibaraki 305, Japan 2National Institute of Agrobiological Resources Tsukuba Science City, Ibaraki 305, Japan

* To whom correspondence should be addressed

Received December 11, 1990. Revised February 5, 1991. Accepted February 5, 1991.

The transgenic tobacco plants have been generated that express the E. coli ß-glucuronldase (GUS) gene under control of the promoter from the rice proliferating cell nuclear antigen (PCNA, DNA polymerase auxiliary protein) gene. GUS expression detected in situ by staining with the chromogenic substrate, 5-bromo-4-chloro-3-indolyl-ß-D-glucuronide (X-Gluc), was restricted to meristems in the organs of the transgenic tobacco plants. This expression responded to the phytohormones which promote callus formation. Furthermore, in situ thymldine uptake showed that the GUS expression pattern corresponded well to the active sites of DNA synthesis. Deletion analysis of the 5' upstream sequence confined the GUS expression pattern to a fragment extending 263 bp upstream of the transcription start site of the rice PCNA gene. Thus, we have identified this fragment as a main regulatory element of the rice PCNA gene promoter.


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