Cloning and expression in E.coli of a synthetic gene for the bacteriocidal protein caltrin/seminalplasmin |
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Authors: | Heaphy Shaun; Singh Mohinder; Gait Michael J |
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Affiliation: | Laboratory of Molecular Biology, Medical Research Council Hills Road, Cambridge CB2 2QH, UK |
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Abstract: | A synthetic gene coding for the bacteriocidal protein caltrin/seminalplasminwas constructed and expressed in Escherichia coli as a fusionwith ß-galactosidase. The gene was designed with arecognition site for the plasma protease, Factor Xa, coded forimmediately prior to the N-terminus of caltrin. The ß-galactosidase-caltrinfusion protein was cleaved with Factor Xa to give caltrin, whichwas identified by its size on SDS-PAGE, its ability to reactwith an antiserum raised to the N-terminal nonapeptide of caltrinand its N-terminal amino acid sequence. After partial purification,synthetic caltrin was found to be active in an assay involvinginhibition of growth of E.coli. |
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Keywords: | caltrin/ seminalplasmin/ synthetic gene/ ß -galactosidase fusion |
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