Polymorphism of the ovine calpastatin gene in some Turkish sheep breeds (original) (raw)
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CALPASTATIN GENE POLYMORPHISM IN ÇINE ÇAPARI AND KARYA SHEEP
This study was carried out to determine Calpastatin gene polymorphism in native Çine Çapari and synthetic Karya sheep in Turkey. Calpastatin is an endogenous inhibitor of calpain. This gene has a key role on meat tenderness after slaughter, and also has been known as candidate gene in muscle growth efficiency. Calpastatin gene was located on 5th chromosome of sheep. Randomly taken blood samples were collected from 97 Çine Çapari and 90 Karya sheep raised in Western Anatolia. Intron I from L domain of the ovine calpastatin gene was amplified by PCR to produce a 565 bp fragment. Then, PCR products were digested with restriction endonuclease enzyme MspI. Digested products were separated by electrophoresis on agarose gel and visualized with gel documentation system. The digestion of the PCR products by MspI enzyme produced fragments of 306 and 259 bp. Data analysis was done using PopGen32 software. In Karya sheep population MM, MN and NN genotypes were identified with 0.296, 0.496 and 0.208 frequencies, M and N allele frequencies were identified with 0.544 and 0.456, respectively. In Çine Çapari sheep population MM, MN and NN genotypes were identified with 0.543, 0,388 and 0.069 frequencies, M and N allele frequencies were identified with 0.737 and 0.26, respectively.
Calpastatin (CAST) gene polymorphism in Kajli, Lohi and Thalli sheep breeds
AFRICAN JOURNAL OF BIOTECHNOLOGY, 2012
important role in the development of muscles and in meat tenderness. The present study was conducted to investigate a calpastatin (CAST) gene polymorphism in Pakistani Thalli, Lohi and Kajli sheep breed. Random blood samples were collected from 300 animals (100 samples from each Thalli, Lohi and Kajli breeds). Genomic DNA was extracted using phenol-chloroform extraction method. A 622 bp CAST gene segment (exon 1C/1D region) was amplified by polymerase chain reaction (PCR) using ovine specific primers. Restriction fragment length polymorphisms (RFLPs) in the amplified fragments were studied using Msp1 restriction enzyme. Frequencies of MM, MN and NN genotypes were found to be 77, 20 and 3% in Lohi breed and 68, 26 and 6% in Kajli breed respectively. In Thalli sheep, only the MM (80%) and MN (20%) genotypes were detected. Chi-Square test (p < 0.05) showed that all the three populations used in this study were in Hardy-Weinberg equilibrium. By comparing the results of this study with those of previous studies, it seems that the MM genotype is the dominant genotype and the M allele is the dominant allele in small ruminant breeds belonging to different geographical locations.
The calpastatin (CAST) gene has a major effect on muscle growth and meat tenderness after slaughter; it is located on the fifth chromosome in sheep. Blood samples were collected from 720 animals in total from Kıvırcık (KIV), Sakız (SZ), Karacabey Merino (KM), and Gökçeada (GA) sheep populations raised in West Anatolia. The PCR products were digested by the restriction endonuclease MspI. Allele frequencies for M and N alleles of the gene were found to be 0.85 and 0.15 in KIV, 0.80 and 0.20 in KM, 0.99 and 0.01 in GA, and 0.34 and 0.66 in SZ sheep, respectively. It was determined that NN genotype frequency was quite lower in KIV (0.04) and KM (0.07) populations than in the SZ breed (0.40). On the other hand, the NN genotype was not observed in the GA population. Populations other than KM were found to be in Hardy-Weinberg equilibrium. In general, allele and genotype frequencies of CAST were similar to those in other studies on different sheep populations. This research is a beginning step for finding candidate genes' effects on meat quantity and quality.
Polymorphism investigation of calpastatin gene in Zel sheep population of Iran by PCR-RFLP method
African Journal of Biotechnology, 2012
Meat tenderness is an important quality characteristic for which consumers are interested. Calpastatin is the calpain inhibitor enzyme and plays an important role in muscle growth and meat quality. The calpastin gene is located on sheep chromosome 5 and its polymorphisms are associated with economic traits. This study was performed to identify calpastatin gene polymorphisms using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Random blood samples were collected from 100 Zel sheep. DNA was extracted by the salting out method and was amplified using PCR. The quantity and quality of the extracted DNA were determined by spectrophotometry and agarose gel electrophoresis. For amplification, a 622 bp fragment exon 1 of the calpastatin gene was used. Amplified fragments were digested by Msp1 restriction enzyme to determine the calpastatin genotypes, and two alleles. M and N were identified. The genotype frequencies of the MM, MN and NN were 0.62, 0.26 and 0.12, respectively in the population under study. Also, the allelic frequency of M and N were 0.75 and 0.25, respectively. Investigation of population genetic structure of Zel sheep revealed that it was not in Hardy-Weinberg equilibrium (p<0.05). The results indicate that it could be useful to consider genetic diversity at calpastatin locus in Zel sheep.
Genotypic Frequency of Calpastatin Gene in Atabi Sheep by PBR Method
2011
Calpastatin have role in regulation muscle growth and meat tenderness after slaughter that its coding gene located on ovine chromosome 5. Studies have shown that this gene is polymorphic in many breeds of sheep and is related with weight gain and carcass traits. In this study blood samples were collected from 120 Atabi sheep located in northern and western Golistan of Iran. Genomic DNA was extracted from blood sample. Gel monitoring and spectrophotometer methods were used to determination quality and quantity of DNA. Exon and entron I from L domain of the ovine calpastatin gene was amplified to produce a 622 bp fragment. The PCR products were digested by restriction endonucleases MspI. Digested products were separated by electrophoresis on 2.5% agarose gel and visualized after staining with ethidium bromide on UV transillumination. The MspI digestion of the PCR products produced digestion fragments of 336 bp and 286 bp. Data analysis was done using PopGen32 software (ver.1.32). In t...
African Journal of Biotechnology, 2014
Calpastatin is a natural occurring inhibitor of calpastatin (CAST) and consequently the balance of calpain-calpastatin activity in muscles is believed to dictate the rate of tenderization in post-mortem meat. Genomic DNA was extracted from 100 sheep blood sample. Polymerase chain reaction was performed to amplify a 622 bp fragment of this gene. Restriction reaction of polymerase chain reaction (PCR) products was done using MspI enzyme. The MspI digestion of the PCR products produced digestion fragments of 336 and 286 bp. The results show that in the population, genotypes AA, AB and BB, respectively, had frequencies 32.2, 63.2 and 4.6, and that this locus was not at Hardy - Weinberg equilibrium in the lori sheep strain (P<0.05). Keywords: Calpastatin gene, polymorphism, lori sheep, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). African Journal of Biotechnology , Vol 13(19), 1952-1954
Genotypic Frequency of Calpastatin Gene in Lori Sheep By PCR-RFLP Method
2011
The effect of Calpains gene polymorphism on the analyzed meat quality traits are discussed in detail in another paper. Calpastatin is a natural occurring inhibitor of calpains and consequently the balance of calpain-calpastatin activity in muscles is believed to dictate the rate of tenderization in postmortem meat. In this study were collected blood samples from 100 Lori sheep. Genomic DNA was extracted from blood sample. Gel monitoring and spectrophotometer methods were used to determination quality and quantity of DNA. MspI enzyme was used for restricting of PCR products. Digested products were separated by electrophoresis on 2% agarose gel and visualized after staining with ethidium bromide on UV transillumination. The PCR product (622 bp) was digested by restriction endonucleases MspI. The MspI digestion of the PCR products produced digestion fragments of 336 bp and 286 bp. Data analysis was done using PopGen32 software (ver.1.32). In the total population of sheep was detected homozygous genotype AA, heterozygous genotype AB and Homozygous genotype BB has been observed for calpastatin gene in Lori sheep strain.
Investigation of Calpastatin Genetic Polymorphism in Egyptian Sheep and Goat Breeds
Biosciences, Biotechnology Research Asia, 2016
Calpastatin is an inhibitor for calpain enzyme system and it has an essential role in meat quality and its tenderness. This trait is of great interest for meat industry and consumers. Molecular genetics techniques help in fishing out of molecular markers which affect the economically production traits in farm animals. Genetic polymorphism of CAST gene and its association with meat quality was reported in different farm animals including cattle, goat and sheep. This work aimed to indentify CAST/MspI genetic polymorphism and its SNPs in Egyptian sheep and goat breeds. The results showed the presence of two genotypes in 140 tested animals, GG and AG with the absence of AA genotypes. The frequencies of GG and AG genotypes in sheep were 65.9% and 34.1%, respectively whereas their frequencies in goat were 56.9% and 43.1%, respectively. The total frequencies for GG and AG genotypes an all 140 tested sheep and goat animals were 62.1% and 37.9%, respectively. The nomenclature of these genotypes was done in this study according to the different nucleotides which were identified after sequencing. The sequence analysis of A and G alleles represented a single nucleotide polymorphism (A→ → → → →G) at position 286 in the amplified fragment. These nucleotide sequences were submitted to GenBank under the accession numbers, KX722533 and KX722534 (Ovis aries, alleles G and A, respectively) and KX722535 and KX722536 (Capra hircus, alleles G and A, respectively). It is concluded that, CAST genetic polymorphisms in Egyptian sheep and goat breeds were similar to those in other sheep populations around the world where the frequencies of alleles and genotypes with G nucleotide is dominant over the others with A nucleotide. Also, A→ → → → →G polymorphism in exon 1 of CAST gene is considered a potential molecular marker for marker assisted selection concerning growth rate where the genotypes with G nucleotide is associated with the significant high weight gain in different small ruminant breeds.
The genotypes for calpastatin (CAST) and calpain (CAPN) loci were determined by PCR-RFLP and PCRSSCP methods. Blood samples were collected from 200 pure-bred Zel sheep from Shirang’s Zel sheep Breeding Station located in south-west of Golestan, Iran. Extraction of genomic DNA was performed based on the modified salting out method. Based on results, two investigated loci were polymorphic and had different gene variants. Heterozygosis was low for both loci. Chi-square test confirmed Hardy-Weinberg equilibrium only in CAST locus using SSCP method. Detected polymorphisms and associations of genetic variation with meat production and tenderness may help to find the effective genotypes of Zel sheep for the economic traits. Keywords: Calpastatin; calpain; molecular methods; polymorphism; sheep
Genetic Variation in the Calpastatin Gene and its Association with Growth Traits in Batur Sheep
KnE Life Sciences
The present study aimed to investigate the association of the CAST genotype and growth traits in Batur sheep. Batur lambs were reared under an intensive feeding system. Bodyweight is measured monthly after weaning until six months of age. Blood representing thirty head were collected, genomic DNA was extracted as samples, and then 200 µl of whole blood samples were used. Specific primers were designed to amplify the CAST gene, samples were sequenced, then the researchers used the BioEdit program to identify any mutation. Calculation of genotypes, gene and allele frequencies, heterozygosities, and Chi-square test was performed. The analysis revealed a total of sixteen polymorphic sites in the CAST coding region. There are four alleles observed (A, G, C, and T), trans-versions at c.92TT loci, and transitions at c.214G>A, c.280G>A, c.301CG. One individual disrupted the reading frame in the whole CAST sequenced. The genotype frequency analysis showed the highest predominance of th...