Cloning, Expression, Characterization, and Tissue Distribution of Cystatin C from Silver Carp (Hypophthalmichthys molitrix)
نویسندگان
چکیده
Cystatins are proteins, which inhibit cysteine proteases, such as papain. In this study, the 336-bp cystatin C gene (family II, HmCysC) of silver carp (Hypophthalmichthys molitrix) was cloned and expressed in Escherichia coli BL21 (DE3). HmCysC encodes mature peptide (HmCystatin C), with 111 amino acids. A typical QXXXG motif found HmCystatin it formed a cluster Cyprinus carpio Danio rerio phylogenetic tree. Quantitative real-time polymerase chain reaction analysis indicated that transcribed at different levels five tested tissues carp. Following purification Ni2+– nitrilotriacetic acid agarose affinity chromatography, displayed molecular weight 20 kDa sodium dodecyl sulfate polyacrylamide gel electrophoresis. Purified had strong inhibitory effects toward proteolytic activity Immunochemical staining anti-HmCystatin antibody showed widely distributed tissues. These results collectively demonstrated properties C, providing information for further studies cystatins from fish organisms.
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ژورنال
عنوان ژورنال: Journal of Agricultural and Food Chemistry
سال: 2021
ISSN: ['1520-5118', '0021-8561']
DOI: https://doi.org/10.1021/acs.jafc.1c00345