Summary
A mutant strain of Wistar rats with L-gulono-γ-lactone oxidase deficiency has recently been established. To investigate this deficiency by DNA and RNA blot hybridization analyses, a fragment of a previously cloned cDNA encoding rat L-gulono-γ-lactone oxidase was used as a probe. When genomic DNA of the mutant rat was digested with several restriction enzymes, the probe hybridized to fragments of the same sizes as those produced from DNA of normal rats. Poly(A)+RNA from the liver of the mutant rat was found to contain an L-gulono-γ-lactone oxidase-specific mRNA of a normal size at a comparable level to that of normal rats. An in vitro translation experiment revealed that the mRNA programmed the synthesis of an enzyme protein which had the same molecular weight as that of the translational product of the normal mRNA, although the amount synthesized was markedly reduced as compared with that synthesized with the normal mRNA. In accordance with this observation, a very low but definite degree of L-gulono-γ-lactone oxidase activity was detected in the microsomes of the mutant rat by a newly developed, highly sensitive method.
Similar content being viewed by others
References
Burns, J. J., Nature180 (1975) 553.
Burns, J. J., Am. J. Med.26 (1959) 740.
Nishikimi, M., and Udenfriend, S., Proc. natl Acad. Sci. USA73 (1976) 2066.
Sato, P., and Udenfriend, S., Archs Biochem. Biophys.187 (1978) 158.
Mizushima, Y., Harauchi, T., Yoshizaki, T., and Makino, S., Experientia40 (1984) 359.
Nishikimi, M., Koshizaka, T., Mochizuki, H., Iwata, H., Makino, S., Hayashi, Y., Ozawa, T., and Yagi, K., Biochem. Int.16 (1988) 615.
Koshizaka, T., Nishikimi, M., Ozawa, T., and Yagi, K., J. biol. Chem.263 (1988) 1619.
Arrand, J. E., in: Nucleic Acid Hybridization. A Practical Approach, p. 17. Eds B. D. Hames and S. J. Higgins. IRL Press, Oxford 1985.
Auffray, C., and Rougeon, F., Eur. J. Biochem.107 (1980) 303.
Maniatis, T., Fritsch, E. F., and Sambrook, J., Molecular Cloning. A Laboratory Manual, p. 197. Cold Spring Harbor Laboratory, Cold Spring Harbor, N.Y. 1982.
McMaster, G. K., and Carmichael, G. G., Proc. natl Acad. Sci. USA74 (1977) 4835.
Koshizaka, T., Nishikimi, M., Tanaka, M., Nakashima, K., Ozawa, T., and Yagi, K., Biochem. Int.15 (1987) 779.
Kiuchi, K., Nishikimi, M., and Yagi, K., Biochemistry21 (1982) 5076.
Kodaka, K., Inagaki, S., Ujiie, T., Ueno, T., and Suda, H., Vitamins59 (1985) 451.
Saheki, T., Imamura, Y., Inoue, I., Miura, S., Mori, M., Ohtake, A., Tatibana, M., Katsumata, N., and Ohno, T., J. inher. metab. Dis.7 (1984) 2.
Wiginton, D. A., Adrian, G. S., Friedman, R. L., Suttle, D. P., and Hutton, J. J., Proc. natl Acad. Sci. USA80 (1983) 7481.
Author information
Authors and Affiliations
Additional information
Acknowledgments. The authors thank Dr Susumu Makino, Shionogi Research Laboratories, Shionogi & Co., Ltd, Japan, for his kind donation of normal (ODS- +/+) and ODS (ODS-od/od) rats. This work was supported in part by Grant-in-Aid (59570103) for Scientific Research from the Ministry of Education, Science and Culture of Japan.
Rights and permissions
About this article
Cite this article
Nishikimi, M., Koshizaka, T., Kondo, K. et al. Expression of the mutant gene for L-gulono-γ-lactone oxidase in scurvy-prone rats. Experientia 45, 126–129 (1989). https://doi.org/10.1007/BF01954844
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01954844