scholarly journals Sequencing the exon 1 of MSTN in Mongolian horse (Equus caballus)

2019 ◽  
Vol 27 (02) ◽  
pp. 3-7
Author(s):  
Sergelen B ◽  
Khaliunaa T ◽  
Myagmarsuren P
Keyword(s):  
Muscle Development ◽  
Equus Caballus ◽  
Reference Sequence ◽  
Amino Acid Difference ◽  
Muscular Tissue ◽  
Long Distance ◽  
Myostatin Gene ◽  
Mstn Gene ◽  
Exon 1 ◽  
Mongolian Horse

Myostatin (MSTN) is a protein responsible for muscular tissue differentiation, development, and growth in mammals. Studies on cattle, mice, pig, and dog have provided sample demonstrations of mutations on myostatin, which strongly affected animal Ankole93200phenotype, particularly muscle development. In horse, myostatin gene is usually studied with respect to racing performance. Most Mongolian native horses do race for long distance and such racing is a very popular sport in Mongolia. On the other hand, the MSTN gene also affects meat yield and quality. We partially sequenced of MSTN gene to explore its polymorphic features in Mongolian horse. A 238 bp long segment of exon 1 was sequenced, using 3130 Genetic Analyser and MEGAX. The sequence was registered with accession number LC216412 in the Gene bank database of NCBI and aligned to four reference sequences (GQ183900.1, KC708233.1, AY840554.2, NM 001081817) from this database. The nucleotide diversity was 0.0084, Tajima test statistic was -1.12. so that there were 233 invariable (monomorphic) sites and 5 variable (polymorphic) sites, in the group of animals compared. The Mongolian horse differed from the reference sequence at 3 sites. There was an amino acid difference in the partial sequence, in which isoleucine in the reference sequence was converted into valine. The results of the study indicate that the Mongolian horse seems to differ substantially from the reference genome of equus caballus. Sequencing of the full length of exon 1 or even the whole MSTN gene, including introns, is strongly advised, to get more insights with regard to the identification and characterization of variants at the myostatin locus in Mongolian horses.

scholarly journals Genetic variation in the first intron and exon of the myostatin gene in several Indonesian cattle populations

2021 ◽  
pp. 1197-1201
Author(s):  
Peni Wahyu Prihandini ◽  
Almira Primasari ◽  
Aryogi Aryogi ◽  
Jauhari Efendy ◽  
Muchamad Luthfi ◽  
...  
Keyword(s):  
Genetic Variation ◽  
Transforming Growth Factor ◽  
Association Studies ◽  
Negative Regulator ◽  
Reaction Products ◽  
Myostatin Gene ◽  
Mstn Gene ◽  
Synonymous Mutations ◽  
Intron 1 ◽  
Exon 1

Background and Aim: Myostatin (MSTN), a member of the transforming growth factor-β family, is a negative regulator of muscle mass. This study aimed to detect the genetic variation of the 1160 bp fragment of exon 1 and part of intron 1 of the MSTN gene in several cattle populations raised in Indonesia. Materials and Methods: Polymerase chain reaction products of the MSTN gene amplified from 92 animals representing 10 cattle populations (Peranakan Ongole [PO], Belgian Blue x PO cross, Rambon, PO x Bali cross, Jabres, Galekan, Sragen, Donggala, Madura, and Bali) were sequenced, compared, and aligned with bovine MSTN of Bos taurus (GenBank Acc. No. AF320998.1) and Bos indicus (GenBank Acc. No. AY794986.1). Results: Four nucleotide substitutions (nt 1045 and 1066 in intron 1; nt 262 and 418 in exon 1) and two indels (nt 807 and 869 in intron 1) were synonymous mutations. Among these substitutions, only the nt 262G>C and nt 418A>G loci were polymorphic in all populations, except Bali cattle. The frequencies of the nt 262C (0.82) and nt 418A (0.65) alleles were highest. For the nt 262G>C locus, the CC genotype had the highest frequency (0.66) followed by GC (0.30) and CC (0.03). For the nt 418A>G locus, the AG genotype had the highest frequency (0.52) followed by AA (0.39) and GG (0.09). Conclusion: The results, showing genetic variations in exon 1 and intron 1 of the MSTN gene, might be helpful for future association studies.

scholarly journals Short Communication: Identification of the exon 1 myostatin gene polymorphism and its association with slaughtered weight in Indonesian Kampung and Broiler Chicken

2020 ◽  
Vol 21 (8) ◽  
Author(s):  
Linda Suhartati ◽  
ISYANA KHAERUNNISA ◽  
ASEP GUNAWAN ◽  
RUKMIASIH RUKMIASIH ◽  
SRI DARWATI ◽  
...  
Keyword(s):  
Gene Polymorphism ◽  
Broiler Chicken ◽  
Muscle Development ◽  
Short Communication ◽  
Negative Regulator ◽  
Silent Mutation ◽  
Muscle Weight ◽  
Myostatin Gene ◽  
Exon 1 ◽  
Meat Type

Abstract. Suhartati L, Khaerunnisa L, Gunawan A, Rukmiasih, Darwati S, Sumantri C, Rizqan. 2020. Short Communication: Identification of the exon 1 myostatin gene polymorphism and its association with slaughtered weight in Indonesian Kampung and Broiler Chicken. Biodiversitas 21: 3893-3897.  Myostatin is widely known as a negative regulator of skeletal muscle development by controlling hypertrophy and hyperplasia. Silent mutation in exon 1 (c.234G>A) was then thought to be associated with chicken growth performances. This study was designed to analyze polymorphisms of c.234G>A mutation of the myostatin gene and its effect on slaughtered weight in Indonesian Kampung and meat-type chicken. The SatI restriction enzyme was used for genotype determination through the PCR-RFLP technique. The effect of genotype on the slaughtered weight was analyzed using the General Linear Model (GLM) procedure. Genotyping was performed on 138 chicken from Kampung (n=67) and meat type (cobb Broiler) (n=71) chicken. The amplification product contained 226 bp nucleotides. This myostatin gene was polymorphic in both chicken population, produced 2 alleles (G and A), and 3 genotypes (GG, GA, and AA). The A allele had a higher frequency than the G allele in all populations. In this study, genotype of AA and GA had higher live weight, carcass weight, breast weight, thigh weight, drumsticks weight, wings weight, breast muscle weight, thighs muscle weight, drumsticks muscle weight than GG genotype of 12 weeks-old Kampung chicken. The results suggested that the c.234G>A mutation in exon 1 could be potentially recommended as a genetic marker for chicken slaughtered weight traits.

MSTN genotype (g.66493737C/T) association with speed indices in Thoroughbred racehorses

2012 ◽  
Vol 112 (1) ◽  
pp. 86-90 ◽  
Author(s):  
Emmeline W. Hill ◽  
Rita G. Fonseca ◽  
Beatrice A. McGivney ◽  
Jingjing Gu ◽  
David E. MacHugh ◽  
...  
Keyword(s):  
Muscle Development ◽  
Mammalian Species ◽  
Maximal Velocity ◽  
Myostatin Gene ◽  
Mstn Gene ◽  
Thoroughbred Racehorses ◽  
Thoroughbred Horses ◽  
Before And After ◽  
Measured Speed ◽  
Dose Dependent

Sequence variation at the equine myostatin gene ( MSTN) locus has previously been shown to have a singular genomic influence on optimum race distance in Thoroughbred racehorses. Myostatin, encoded by the MSTN gene, is a member of the TGF-β superfamily that regulates skeletal muscle development in a range of mammalian species including the horse. In the Thoroughbred, the C-allele at the g.66493737C/T SNP has been found at significantly higher frequency in subgroups of the population that are suited to fast, short distance, sprint races and also influences body composition phenotypes. We investigated the influence of the g.66493737C/T SNP on speed indexes measured in a cohort of n = 85 Thoroughbred horses-in-training. We found significant associations between genotypes at the g.66493737C/T SNP and all measured speed variables: Dist6 [distance travelled during 6 s before and after maximal velocity (Vmax); P = 0.0040], Vmaxt (duration at Vmax; P = 0.0249), Vmax ( P = 0.0265), Dist6b (distance travelled during 6 s before Vmax; P = 0.0032), and Dist6a(distance travelled during 6 s after Vmax; P = 0.0317). For each measure, horses with the C/C and C/T genotypes outperformed T/T horses, indicating the requirement for at least one C-allele to improve speed. For the most significantly associated variables (Dist6 and Dist6b) the C/C cohort performed better than the T/T cohort with the heterozygotes intermediate, indicating a dose-dependent manifestation. These findings clearly indicate that variation at the MSTN gene influences speed in Thoroughbred horses.

Effect of kinin B2 receptor ablation on skeletal muscle development and myostatin gene expression

Neuropeptides ◽  
2010 ◽  
Vol 44 (2) ◽  
pp. 209-214 ◽  
Author(s):  
K. de Picoli Souza ◽  
E.C. Batista ◽  
E.D. Silva ◽  
F.C. Reis ◽  
S.M.A. Silva ◽  
...  
Keyword(s):  
Gene Expression ◽  
Skeletal Muscle ◽  
Muscle Development ◽  
Skeletal Muscle Development ◽  
Myostatin Gene

94 Myostatin gene editing by CRISPR/Cas9 technology of Brangus fetal fibroblasts to produce edited embryos by cloning

2021 ◽  
Vol 33 (2) ◽  
pp. 154
Author(s):  
J. I. Bastón ◽  
D. Viale ◽  
M. Olguin ◽  
E. Wiedenmann ◽  
V. Arnold ◽  
...  
Keyword(s):  
Gene Expression ◽  
Production Efficiency ◽  
Gene Editing ◽  
Premature Stop Codon ◽  
Exogenous Dna ◽  
Exon 2 ◽  
Myostatin Gene ◽  
Mstn Gene ◽  
Cattle Breeds ◽  
Fetal Fibroblasts

Some European cattle breeds, such as Charolais and Maine Anjou, have natural mutations in the myostatin gene (MSTN) that inhibit its expression and result in an increase in muscle mass and protein content. An innovative hallmark would be the invitro introduction of this genotype in South America cattle breeds to improve their commercial value. To achieve this, we aimed to disrupt MSTN gene expression in bovine fetal fibroblasts (BFFs) using CRISPR/Cas9 technology and to generate MSTN-edited embryos by somatic cell nuclear transfer (SCNT). BFFs were isolated from a cloned fetus of a Brangus bull with a prized genetic background and nucleofected with the Cas9 ribonucleoprotein-gRNA complex previously assessed to target exon 2 of the bovine MSTN gene. To evaluate MSTN editing, genomic DNA from the wild-type (WT) and nucleofected BFFs were isolated, exon 2 of MSTN was amplified by PCR, and the PCR product was Sanger sequenced. In all cases, the sequencing results were analysed using the indel Synthego software tool. According to indel analysis, MSTN gene editing efficiency of the BFFs was 58.83%±3.2 (n=6). The resulting edit consisted of insertion of thymine in exon 2 of the MSTN gene that shifted the gene reading frame, introducing a premature stop codon and generating a truncated MSTN protein. The nucleofected BFFs were then used to generate embryos by SCNT, and WT BFFs were included as controls. Embryo cleavage and blastocyst development rates were evaluated at Day 2 and 7, respectively (Chi-squared test, P<0.05). Although lower cleavage rates were obtained in the MSTN-edited BFFs group [65.3% (n=273/418) vs. 87.6% (n=169/193)], no differences were observed at the blastocyst stage [19.1% (n=80/418) vs. 25.4% (n=49/193)]. To confirm the efficiency of MSTN editing in the cloned embryos, 10 blastocysts generated with the MSTN-edited BFFs were individually analysed for MSTN exon 2 sequence as described before. The results showed that 30% of the blastocysts (3/10) presented a homozygous biallelic edition, which consisted of a thymine base insertion, as expected. In summary, the strategy we used allowed production of MSTN null cloned Brangus embryos avoiding putative undesired integration of exogenous DNA into the bovine genome, such as plasmid sequence, regardless of off-target occurrences. We conclude that CRISPR/Cas9 is an efficient technique to disrupts MSTN gene expression in bovine embryos; this work represents a step toward improving the production efficiency of South American cattle breeds.

Genetic Diversity Identification and Haplotype Distribution of Myostatin Gene in Goats

2020 ◽  
Author(s):  
Cheng-Li Liu ◽  
Ri-Su Na ◽  
Wei-Wei Ni ◽  
Guang-Xin E ◽  
Yan-Guo Han ◽  
...  
Keyword(s):  
Genetic Diversity ◽  
Muscle Development ◽  
Haplotype Frequency ◽  
Negative Regulator ◽  
Myostatin Gene ◽  
Polymorphic Loci ◽  
Boer Goat ◽  
Haplotype Distribution ◽  
Meat Goats ◽  
Significant Difference

Background: Myostatin (MSTN) is a highly conserved protein that acts as a negative regulator of skeletal muscle growth. MSTN gene is closely associated with multiple biological functions and its mutations are directly linked to muscle development in different species. Loss of MSTN functionality causes the phenotype to appear in the form of ‘double musculature’, among others in cattle, sheep and house mice.Methods: The mixed DNA pool of Boer and Dazu black goats to sequence MSTN coding and noncoding regions. Snapshot typing technology was used to analyse three goat production types, namely, Boer (meat goats), Nubian (breast and meat dual-use goat), Dazu black (local breeds) and Youzhou black goats (local breeds). Polymorphic loci in MSTN were used from four goat populations to construct haplotypes and calculate haplotype frequency distribution through NETWORK and MEGA to construct a phylogenetic tree and visualize phylogenetic relationship.Result: From these populations, 18 haplotypes were constructed using 20 polymorphic loci. Phylogenetic analysis revealed a significant difference in the haplotype distribution of MSTN in different goat production types. The 18 haplotypes were divided into two clusters. The haplotypes carried by the Boer goat belonged to Cluster I and those carried by the Nubian goat and two local goat breeds belonged to both clusters. Chinese local goats and complex production goats carried more haplotypes of MSTN and had a richer genetic diversity than other production types did. Moreover, local and complex production goats had high-frequency haplotypes of the MSTN of meat goats and had high potential for breeding.

scholarly journals New possibilities of effective breeding in cattle based on the study of the genome

2017 ◽  
Vol 19 (79) ◽  
Author(s):  
V. Y. Bodnaruk ◽  
L. I. Muzyka ◽  
P. V. Bodnar ◽  
A.J. Zhmur ◽  
T. V. Orihivsjkyj
Keyword(s):  
Growth Hormone ◽  
Genetic Markers ◽  
Quantitative Traits ◽  
Muscle Development ◽  
Single Gene ◽  
Molecular Genetic ◽  
Chromosome 2 ◽  
Myostatin Gene ◽  
Molecular Genetic Markers ◽  
Breeding Work

The article contains generalized literary data on the results of genome research based on molecular genetic methods in connection with the productive qualities of cattle that can be used to accelerate and improve breeding work. The study of the human genome has given impetus to the development of medicine, biotechnology and pharmacogenetics. Similarly, new research on the genome of cattle gives qualitatively different possibilities for using these data in the selection and production of agricultural products, as well as in controlling its quality. Molecular genetic markers inform about the polymorphism of genes and allow to detect individual genes and gene complexes that carry information about a certain feature. Based on such studies, gene pool can be formed with a certain combination. An alternative way of molecular-genetic marking of performance is to study the polymorphism of structural genes, allelic variants which are directly related to the desired phenotypic manifestation, namely: kappa-casein (CSN3), veta-lactaglobulin (BLG), somatotropin (GH), and myostatin (MSTN). Modern breeding work with cattle is associated with the establishment of a connection between the polygenic signs of productivity and the «main» genes of quantitative traits, the polymorphism of which affects the final output of the protein product. As candidate genes that affect lactation productivity in cattle, first of all the genes of milk proteins, in particular kappa-casein, are examined. The gene for the somatotropic hormone (GH), a growth hormone in cattle, is a polypeptide consisting of 191 amino acids and is encoded by a single gene, which is localized in 19 chromosomes. Growth hormone plays a key role in stimulating the synthesis of protein, cell division, and body growth. Myostatin – one of the regulators of skeletal muscle development is the myostatin gene, which refers to a family of transforming growth factors. The gene of myostatin in the Bovine species is localized in chromosome 2 and carries the muscle hypertrophy locus, there is also a homologous fragment of human chromosome 2, where the locus of this gene is limited. The presence of the gene of myostatin, as one of the locus of quantitative traits of beef, can be used as a marker for genetic mapping. After discovering mutations in the gene of the myostatin, they came to the conclusion, that it is not the only gene that controls the growth and muscle mass of animals. Molecular genetic markers allow you to receive information about the polymorphism of genes and to identify individual genes and gene «ensembles» that carry the desired complex of features.

scholarly journals Myostatin (MSTN) Gene Indel Variation and Its Associations with Body Traits in Shaanbei White Cashmere Goat

Animals ◽  
2020 ◽  
Vol 10 (1) ◽  
pp. 168 ◽  
Author(s):  
Yi Bi ◽  
Bo Feng ◽  
Zhen Wang ◽  
Haijing Zhu ◽  
Lei Qu ◽  
...  
Keyword(s):  
Transforming Growth Factor Beta ◽  
Muscle Development ◽  
Transforming Growth Factor ◽  
Growth Traits ◽  
Body Height ◽  
Cashmere Goat ◽  
Sample Sizes ◽  
Association Analyses ◽  
Mstn Gene ◽  
Almost All

Myostatin (MSTN) gene, also known as growth differentiation factor 8 (GDF8), is a member of the transforming growth factor-beta super-family and plays a negative role in muscle development. It acts as key points during pre- and post-natal life of amniotes that ultimately determine the overall muscle mass of animals. There are several studies that concentrate on the effect of a 5 bp insertion/deletion (indel) within the 5’ untranslated region (5’ UTR) of goat MSTN gene in goats. However, almost all sample sizes were below 150 individuals. Only in Boer goats, the sample sizes reached 482. Hence, whether the 5 bp indel was still associated with the growth traits of goats in large sample sizes which were more reliable is not clear. To find an effective and dependable DNA marker for goat rearing, we first enlarged the sample sizes (n = 1074, Shaanbei White Cashmere goat) which would enhance the robustness of the analysis and did the association analyses between the 5 bp indel and growth traits. Results uncovered that the 5 bp indel was significantly related to body height, height at hip cross, and chest width index (p < 0.05). In addition, individuals with DD genotype had a superior growing performance than those with the ID genotype. These findings suggested that the 5 bp indel in MSTN gene are significantly associated with growth traits and the specific genotype might be promising for maker-assisted selection (MAS) of goats.

Myostatin silencing effect on basic helix–loop–helix transcription factors in caprine foetal fibroblast cells

2017 ◽  
Author(s):  
Biswajyoti Borah ◽  
Ajit Pratap Singh ◽  
Hamen Gogoi ◽  
Amlan Jyoti Phukan ◽  
Bikash Chandra Sarkhel
Keyword(s):  
Transcription Factors ◽  
Muscle Development ◽  
Pearson Correlation ◽  
Negative Regulator ◽  
Fibroblast Cells ◽  
Animal Cells ◽  
Myostatin Gene ◽  
Basic Helix Loop Helix ◽  
Helix Loop Helix

Transgenic food animal production is one of the potential and need oriented research to mitigate the food crises of the world. In vitro gene silenced animal cells and making use of these cells for transgenesis one of the suitable way to produce productive animals. Myostatin is a negative regulator of muscle growth, has the potential to increase the muscle mass upon its silencing. Four Hush 29-mer anti- myostatin (MSTN) shRNA constructs were checked for myostatin gene silencing in caprine foetal fibroblast cells and its subsequent effect on basic helix– loop–helix (bHLH) transcription factors. These factors are necessary for the terminal differentiation, proliferation, and homeostasis of muscle development. Different shRNA constructs displayed 55.1 to 91.5% (p less than 0.01) of myostatin silencing in caprine foetal fibroblast cells and upregulation of myogenic gene. Upregulation of 7.97 to 111.67 % for MyoD, 77.0 % to 319.47 % for myogenin, 16.67 % to 138.0 % for Myf5 were observed . The Pearson correlation established a negative correlation between myostatin and genes under study. Result suggests that knockdown of MSTN a potential approach to improve caprine musculatures.

scholarly journals Molecular Characterization and Phylogeny Based Analysis of Intron I Sequence of Myostatin (MSTN) Gene in Iranian Makuei Sheep Breed

2016 ◽  
Vol 16 (4) ◽  
pp. 1007-1018
Author(s):  
Mohammad Farhadian ◽  
Ali Hashemi
Keyword(s):  
Muscle Development ◽  
Negative Regulator ◽  
Nucleotide Sequencing ◽  
Meat Production ◽  
Potential Candidate ◽  
Nucleotide Polymorphisms ◽  
Single Strand Conformational Polymorphism ◽  
Mstn Gene ◽  
Potential Candidate Gene ◽  
Intron 1

Abstract Myostatin (MSTN), a negative regulator of skeletal muscle development, acts as a potential candidate gene used to increase muscle mass. Likewise, sheep MSTN gene has an important role in meat production. MSTN is made up of 376 amino acids, and is synthesized as a precursor protein. To investigate the MSTN in Iranian native Makuei sheep, a polymerase chain reaction-single strand conformational polymorphism (PCR-SSCP) analysis was used. Genomic DNA was isolated from the blood samples. A 417-bp of MSTN intron I segment was amplified using locus-specific primers. Four SSCP patterns were identified and nucleotide sequencing of the Makuei sheep MSTN gene was done and registered in the NCBI GenBank with “KJ526625” number. Three novel single nucleotide polymorphisms (SNPs) were found in the samples. These SNPs are found in 224bp, 226bp and 242bp locations. Accordingly three substitutions (c.224C>T; c.226A>G; c.242G>T) were observed in the intron 1 region of MSTN gene. The effects of the observed SNPs on breeding values of some biometric traits were investigated and the substitution of c.226A>G was found to be associated with heart girth (HG) and leg circumference (LC). Phylogenetic analysis, based on the nucleotide sequences indicated similar evaluation with the GenBank reference sequences. It seems that the observed polymorphisms of the ovine MSTN gene are associated with HG and LC traits.

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