Analysis of sequence variations of Calpastatin gene of inhibitory region in Cyprinus carpio var. communis
Journal Title: JOURNAL OF ADVANCES IN BIOTECHNOLOGY - Year 2014, Vol 4, Issue 3
Abstract
Calpastatin gene, an endogenous inhibitor of calpain enzyme, is important for maintaining muscle texture and postmortem degradation of myofibrils. Present study was carried out for the identification of SNPs in the inhibitory region of calpastatin gene in common carp Cyprinus carpio var. communis for genetic improvement of genotypes. Primers were designed from cDNA sequences of Atlantic salmon (AFV08638.1), Rainbow trout (NP_001118010.1) and cDNA as well as genomic DNA sequences of Zebra fish (NM_001130591.2) to amplify the CAST gene in Common carps. Four exons (13, 14, 19 and 20) and intervening introns demarcated by zebrafish genomic data base of 316 and 311 base pairs were amplified. The sequencing and insilico translation was done for the identification and characterization calpastatin. The results showed that Cyprinus carpio var. communis have two highly conserved sequence motifs DTLPP, GYR repeats in the inhibitory domain which corresponds to the exon-14 and exon-20 of Zebra fish. The single amino acid change in the inhibitory motif G (GGT)/X (X = Q 394aa (CAA) in Danio rio, Sequence ID: ref|NP_001124063.2, E 491aa (GAG) in Salmo salar, Sequence ID: ref|NP_001167162.1, K 223aa (AAG) in Onchorhyncus mykiss long isoform, Sequence ID: ref|NP_001118010.1, and E 54aa (GAG) in Onchorhyncus mykiss short isoform, Sequence ID: ref|NP_001118117.1) have role in the diversification of inhibitory activity of the protein due to species diversification. Another interesting characteristic feature of the inhibitory motif of exon-20 in Common carps having Glysine (GGT) amino acid and rest instead of Glutamic acid (GAG) at 548th aa in Danio rio and 689th aa Salmo salar. It was also observed that Common carp shear (94-95%) CAST gene sequence with Zebra fish while as only (52-54%) sequence homology was observed with Atlantic salmon and Rainbow trout due to species divergence. Phylogenetic analyses of different species revealed telosts are more primitive than tetrapods. Variations observed in inhibitory domain play important role in postmortem proteolysis. Development of markers for meat tenderness and fillet quality based on the mutations in the inhibitory region will help in selecting genotypes of better fillet/texture quality.
Authors and Affiliations
Dr Mohammed Iqbal Mir, Salma Khan, F A Shah, Gh Nabi Lone, Masood H Balkhi
Keywords
Related Articles
- EP ID EP653877
- DOI 10.24297/jbt.v4i3.4997
- Views 64
- Downloads 0
Acute and Sub-chronic (28-day) intraperitoneal toxicity studies of the Methanol Root Extract of Securidaca longepedunculata Fresen (Polygalaceae) in Rats.
Despite the wide spread use of Securidaca longepedunculata in all parts of Africa, not much has been reported in the literature about its toxicity. The few reports available are in connection with its trypanocidal and in...
Molecular Typing by Polymerase Chain Reaction Sequence Specific Primers (PCR- SSP) of Human Leukocyte Class I and Class II Alleles in a Sample of Iraqi Visceral Leishmaniasis Patients
Objectives: This study aimed to investigate the association between HLA alleles and visceral leishmaniasis (VL) in a sample of Iraqi patients. Methods: A total of 30 patients were studied, in addition to 20 age, gender...
Evaluation of Candida guilliermondii cell growth and one-pot fermentation for xylitol bioproduction
Xylitol is a polyalcohol with a high sweetener power that presents healthy benefits when compared to sucrose, such as prevention of caries and metabolism only partially dependent on insulin. Its bioproduction, besides be...
A Carbohydrate Hydrolysis Enzymes encoding Genes in Neurospora crassa
Neurospora crassa, NCU05882.7 (423aa) and NCU09774.7 (303 aa) (NCU, Neurospora 7 crassa unit) genes encoding a Cellulase, which hydrolysis the Cellulose. In addition to that, 8 reporting here other 35 Carbohydrate hydrol...
Development, Optimisation, Validation and Inter-Laboratory Verification of a Reversed Phase HPLC Method for Quantification of Human Recombinant Insulin
HPLC methods for insulin in official monographs require extended runtimes and elevated temperatures. Inter-laboratory reproducibility of HPLC methods obtained from published literature is an on-going challenge, moreso fo...