Expression of maize prolamins in Escherichia coli |
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Authors: | Shu-Zhen Wang Asim Esen |
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Affiliation: | Department of Biology, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, U.S.A. |
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Abstract: | We have constructed a cDNA expression library of developing corn (Zea mays L.) endosperm using plasmid pUC8 as vector and Escherichia coli strain DH1 as host. The expression library was screened with non-radioactive immunological probes to detect the expression of gamma-zein and alpha-zein. When anti-gamma-zein antibody was used as the probe, 23 colonies gave positive reactions. The lengths of cDNA inserts of the 23 colonies were found to be 250–900 base pairs. When anti-alpha zein antibody was used, however, fewer colonies gave positive reactions. The library was also screened by colony-hybridization with 32P-labeled DNA probes. Based on immunological and hybridization screening of the library and other evidence, we conclude that alpha-zein was either toxic to E. coli cells or rapidly degraded whereas gamma-zein and its fragments were readily expressed. |
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Keywords: | zein expression AMV avian myeloblastosis virus KD kilodalton MW molecular weight SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis |
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