PUBLICATION
Cloning, Expression, Characterization, and Tissue Distribution of Cystatin C from Silver Carp (Hypophthalmichthys molitrix)
- Authors
- Li, R., Tan, X., Li, S., Jin, Y., Li, S., Li, S., Takala, T.M., Saris, P.E.J.
- ID
- ZDB-PUB-210422-4
- Date
- 2021
- Source
- Journal of Agricultural and Food Chemistry 69(17): 5144-5154 (Journal)
- Registered Authors
- Keywords
- cloning, cystatin C, protein quantification, silver carp (Hypophthalmichthys molitrix), tissue distribution, transcription level
- MeSH Terms
-
- Carps*/genetics
- Tissue Distribution
- Cystatins*/genetics
- Animals
- Cystatin C
- Phylogeny
- Cloning, Molecular
- PubMed
- 33881846 Full text @ J. Agric. Food Chem.
Citation
Li, R., Tan, X., Li, S., Jin, Y., Li, S., Li, S., Takala, T.M., Saris, P.E.J. (2021) Cloning, Expression, Characterization, and Tissue Distribution of Cystatin C from Silver Carp (Hypophthalmichthys molitrix). Journal of Agricultural and Food Chemistry. 69(17):5144-5154.
Abstract
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 the mature peptide of cystatin C (HmCystatin C), with 111 amino acids. A typical QXXXG motif was found in HmCystatin C and it formed a cluster with Cyprinus carpio and Danio rerio cystatin C in the phylogenetic tree. Quantitative real-time polymerase chain reaction analysis indicated that HmCysC was transcribed at different levels in five tested tissues of silver carp. Following purification with Ni2+- nitrilotriacetic acid agarose affinity chromatography, HmCystatin C displayed a molecular weight of 20 kDa in sodium dodecyl sulfate polyacrylamide gel electrophoresis. Purified HmCystatin C had strong inhibitory effects toward the proteolytic activity of papain. Immunochemical staining with anti-HmCystatin C antibody showed that HmCystatin C was widely distributed in silver carp tissues. These results collectively demonstrated the properties of HmCystatin C, providing information for further studies of cystatins from fish organisms.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping