Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

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Hokari, S. et al.

Hokari, Miura, Koyama, Kobayashi, Komine, Komoda,

A restriction endonuclease assay for expression of human alpha-amylase isozymes.

BACKGROUND: The alpha-amylase isozymes can be detected separately by electrophoresis; however, sometimes the identification is difficult because of their microheterogeneity. In the present study, we tried to establish a convenient method for the detection of alpha-amylase isozyme expression. METHODS: The procedure is based on three different restriction sites presented in those genes; a PstI site in both AMY 2A and 2B genes, a HaeII site in both AMY 1 and 2A genes, and a BamHI site in AMY 2B gene. After amplification from total tissue RNAs by RT-PCR with primers that were able to cover each exon, the products were cleaved with corresponding restriction endonucleases. RESULTS: This method was applied to human samples from the parotid gland, liver (non-hepatoma), hepatoma and white blood cells (WBCs). The results indicated that the parotid gland and hepatoma (also liver) clearly expressed AMY 1 and AMY 2B genes, respectively. However, AMY 2B gene was also expressed apparently in WBCs, which produced salivary-type isozyme of the alpha-amylase, although the amylase protein was not able to identify for the hepatic isozyme. CONCLUSIONS: The method presented here might be convenient and useful for the determination of alpha-amylase isozyme expression in humans.[1]

References

A restriction endonuclease assay for expression of human alpha-amylase isozymes. Hokari, S., Miura, K., Koyama, I., Kobayashi, M., Komine, S., Komoda, T. Clin. Chim. Acta (2002) [Pubmed]

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