Synthesis and expression of a gene coding for the calcium-modulated protein S100 beta and designed for cassette-based, site-directed mutagenesis.
作者： L J Van Eldik , J L Staecker , F Winningham-Major
DOI: 10.1016/S0021-9258(18)68573-4
关键词:
摘要: As an initial step in studies aimed at addressing the question of what common and unique features S100 family proteins are related to their specific functions localizations, a gene coding for one proteins, beta, has been prepared by ligation 12 overlapping, synthetic oligonucleotides. Automated DNA sequence analysis demonstrated that final construct expected structure. The was inserted into plasmid vector contains tac promoter ampicillin-resistance gene, thus allowing both amplification direct expression cloning Escherichia coli. designed allow rapid, efficient changes single or multiple amino acids using cassette-based mutagenesis while is resident vector. expressed protein (VUSB-1) indistinguishable from bovine brain beta terms electrophoretic mobility, reactivity with antibodies acid composition, partial analysis. Preparations also functionally similar as determined aldolase activator activity neurite extension factor activity, supporting concept these activities property polypeptide.
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