scholarly journals Detection of F94L mutation of the MSTN gene in four Indonesian local cattle breeds

2020 ◽  
Vol 45 (1) ◽  
pp. 7-14
Author(s):  
S. Anwar ◽  
S. D. Volkandari ◽  
A. S. Wulandari ◽  
W. P. B. Putra ◽  
E. Sophian ◽  
...  
Keyword(s):  
Meat Quality ◽  
Wild Type ◽  
Myostatin Gene ◽  
Mstn Gene ◽  
Cattle Breeds ◽  
Holstein Friesian ◽  
Pcr Rflp ◽  
Genotype Identification ◽  
The Status ◽  
Type C

The F94L mutation of the MSTN gene (MSTN-F94L) is considered not to cause disrupted the function of the myostatin gene drastically. Interestingly, this mutation has a very significant effect on muscle mass, carcass, or meat yield and meat quality without any associated severe negative problems. This study aimed to confirm the MSTN-F94L mutation in four local cattle breeds in Indonesia. A total of 518 individuals (140 of Bali, 107 of Sumbawa, 168 of Pasundan, and 103 of Holstein-Friesian (H-F) cattle) were used in this study. Genotype identification was performed by PCR-RFLP method. In the present study showed that the wild-type C allele was fixed (1.000) in Bali, Sumbawa, and HF cattle. However, the wild-type C allele and the mutant A allele were found in Pasundan cattle, even though the frequency of the mutant A allele was very low (0.012). Therefore, in conclusion, the mutation of the MSTN-F94L was detected in Pasundan cattle but no in all three cattle breeds. However, the presence of the mutant A allele in Pasundan cattle allegedly derived from Limousin bulls. The further investigation in other local and exotic breeds and its crossing will answer the status of the MSTN-F94L mutation in local cattle breeds in Indonesia.

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.

scholarly journals Genetic Polymorphism in a Selective Intron of Ovine Myostatin Gene and Its Putative Relation with Carcass Traits in Sheep

2020 ◽  
Author(s):  
M. Metta ◽  
D. V. Praneeth ◽  
R. Vinoo ◽  
K. Sudhakar ◽  
S. Jagadeswararao
Keyword(s):  
Meat Quality ◽  
Polymorphic Locus ◽  
Association Studies ◽  
Donor Site ◽  
Snp Markers ◽  
Limited Data ◽  
Myostatin Gene ◽  
Genetic Groups ◽  
Pcr Rflp ◽  
Representative Samples

The main aim of the present study is to identify a suitable polymorphic locus in the myostatin gene of sheep that could be associated with production in local sheep genetic groups of Andhra Pradesh province in India. Representative samples from three local genetic groups were used in the present study namely Nellore Jodipi, Nellore Brown and Macherla Brown. Two PCR-RFLP based SNP markers located in GDF-8 (myostatin-MYST) locus were used in the present study. A PCR-RFLP assay was developed for a SNP located in the intron1 (rs119102825) of the myostatin gene. The second marker is located in exon3 which was obtained from previous studies. The SNP located in Intron1 is polymorphic and the SNP located in exon3 is monomorphic. The polymorphism information content (PIC) of the SNP in intron1 is 0.37. Association studies with limited data showed lack of association of genotypes with body weight at different ages. However, based on bioinformatic prediction, it is likely that the SNP in the intron1 locus may be involved in meat quality determination as it is close to the donor site of intron and the variation has potentiality for enhancement or repression of the gene expression. Further association studies with meat quality traits would help in understanding functional implications of the polymorphism.

scholarly journals Variants in the Myostatin Gene and Physical Performance Phenotype of Elite Athletes

Genes ◽  
2021 ◽  
Vol 12 (5) ◽  
pp. 757
Author(s):  
Valentina Ginevičienė ◽  
Audronė Jakaitienė ◽  
Erinija Pranckevičienė ◽  
Kazys Milašius ◽  
Algirdas Utkus
Keyword(s):  
Physical Performance ◽  
Elite Athletes ◽  
Performance Status ◽  
Muscle Growth ◽  
Endurance Performance ◽  
Negative Regulator ◽  
Myostatin Gene ◽  
Mstn Gene ◽  
Homozygous Genotype ◽  
The Status

The MSTN gene is a negative regulator of muscle growth that is attracting attention as a candidate gene for physical performance traits. We hypothesised that variants of MSTN might be associated with the status of elite athlete. We therefore sought to study the potential role of MSTN in the physical performance of athletes by analysing the whole coding sequence of the MSTN gene in a cohort of Lithuanian elite athletes (n = 103) and non-athletes (n = 127). Consequently, two genetic variants were identified: the deletion of one of three adenines in the first intron (c.373+90delA, rs11333758) and a non-synonymous variant in the second exon (c.458A>G, p.Lys(K)153Arg(R), rs1805086). Among all samples, the MSTN rs1805086 Lys(K) allele was the most common form in both groups. Homozygous genotype for the less common Arg(R) allele was identified in only one elite canoe rower, and we could find no direct association between rs1805086 and successful results in elite athletes. Surprisingly, the intronic variant (rs11333758) was abundant among all samples. The main finding was that endurance-oriented athletes had 2.1 greater odds of being MSTN deletion genotype than non-athletes (13.6% vs. 0.8%). The present study confirms the association of the polymorphism rs11333758 with endurance performance status in Lithuanian elite athletes.

scholarly journals Polymorphism of 5’UTR myostatin gene indel (g.1256/TTTTA) and its association with body weight in Boerka crossbred goat

2020 ◽  
Vol 45 (3) ◽  
pp. 163-172
Author(s):  
R. Ismail ◽  
E. Handiwirawan ◽  
S. Elieser ◽  
J. Jakaria
Keyword(s):  
Body Weight ◽  
Birth Weight ◽  
Linear Model ◽  
Restriction Enzyme ◽  
Direct Sequencing ◽  
General Linear Model ◽  
Myostatin Gene ◽  
Mstn Gene ◽  
Genotype Frequencies ◽  
Pcr Rflp

This study aimed to identify the variation of 5’UTR Myostatin (MSTN) gene and its association to body weight in Boer, Kacang, and Boerka goats. DNA samples were obtained from 149 heads of the goats from the Indonesian Goat Research Center, Sungei Putih, North Sumatera. Polymorphism identification was conducted by direct sequencing and PCR-RFLP with DraI as the restriction enzyme for indel g.1256/TTTTA. Analysis of variance for body weight was carried out using the General Linear Model (GLM). The 5’UTR MSTN gene|DraI was monomorphic in Kacang but polymorphic in Boer and Boerka. Genotype frequencies for Boer 0.40(AA), 0.43(AB), 0.17(BB); Kacang 1.00(AA); Boerka 0.53(AA), 0.37(AB), 0.10(BB). The allele frequencies for Boer 0.62(A), 0.38(B); Kacang 1.00(A); Boerka 0.72(A), 0.28(B), respectively. AB was the most frequent genotype among Boer, but AA was the most frequent in Kacang and Boerka. Indel g.1256/TTTTA has a significant effect (P<0.05) only on birth weight (BW) of Boer, but no significant effect on other bodyweight parameters both in Boer and Boerka.AA genotype has the highest BW (P<0.05) than AB, but it’s not significantly different from BB. Indel g.1256/TTTTA could be used as a genetic marker for the birth weight of Boer but not in Boerka goats. 

scholarly journals PCR-based genotyping assays to detect germline APC variant associated with hereditary gastrointestinal polyposis in Jack Russell terriers

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Kyoko Yoshizaki ◽  
Akihiro Hirata ◽  
Hiroyuki Matsushita ◽  
Naohito Nishii ◽  
Mifumi Kawabe ◽  
...  
Keyword(s):  
Mutant Allele ◽  
False Negative ◽  
Disease Onset ◽  
Pcr Amplification ◽  
Buccal Swab ◽  
Wild Type ◽  
Gastrointestinal Polyposis ◽  
Pcr Rflp ◽  
Formalin Fixed Paraffin Embedded ◽  
Rflp Assay

Abstract Background The prevalence of gastrointestinal (GI) neoplastic polyps in Jack Russell terriers (JRTs) has increased in Japan since the late 2000s. Recently, we demonstrated that JRTs with GI polyps harbor identical germline variant in the APC gene (c.[462_463delinsTT]) in the heterozygous state. Thus, this disease is an autosomal dominant hereditary disorder. Although the affected JRTs have distinct features, such as the development of multiple GI polyps and an early age of disease onset, genetic testing is indispensable for a definitive diagnosis. Here, polymerase chain reaction (PCR)-based assays capable of detecting germline APC variant were designed and validated using synthetic wild-type and mutant DNAs and genomic DNAs from carrier and non-carrier dogs. Result First, the PCR-restriction fragment length polymorphism (PCR-RFLP) assay was developed by taking advantage of the germline APC variant creating a new restriction site for MseI. In the PCR-RFLP assay, the 156-bp region containing the variant site was amplified by PCR and subsequently digested with MseI, yielding diagnostic 51 and 58 bp fragments from the mutant allele and allowing determination of the APC genotypes. It was possible to determine the genotypes using genomic DNA extracted from the peripheral blood, buccal swab, or formalin-fixed paraffin-embedded tissue. Next, a TaqMan duplex real-time PCR assay was developed, where a 78-bp region flanking the variant was amplified in the presence of wild-type allele- and mutant allele-specific fluorescent probes. Using blood-derived DNA, altogether 40 cycles of PCR amplification determined the APC genotypes of all examined samples by measuring the fluorescence intensities. Importantly, false-positive and false-negative errors were never detected in both assays. Conclusion In this study, we developed highly reliable genetic tests for hereditary GI polyposis in JRTs, providing accurate assessment of the presence of the causative germline APC variant. The genotyping assays could contribute to the diagnosis and prevention of hereditary GI polyposis in dogs.

scholarly journals Associations between the Bovine Myostatin Gene and Milk Fatty Acid Composition in New Zealand Holstein-Friesian × Jersey-Cross Cows

Animals ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1447
Author(s):  
Ishaku L. Haruna ◽  
Yunhai Li ◽  
Ugonna J. Ekegbu ◽  
Hamed Amirpour-Najafabadi ◽  
Huitong Zhou ◽  
...  
Keyword(s):  
Fatty Acid ◽  
New Zealand ◽  
Nucleotide Sequence ◽  
Milk Fat ◽  
Linear Mixed Effect Model ◽  
Polymorphism Analysis ◽  
Mixed Effect ◽  
Myostatin Gene ◽  
Holstein Friesian ◽  
Nucleotide Sequence Variation

The myostatin gene (MSTN), which encodes the protein myostatin, is pleiotropic, and its expression has been associated with both increased and decreased adipogenesis and increased skeletal muscle mass in animals. In this study, the polymerase chain reaction, coupled with single strand conformation polymorphism analysis, was utilized to reveal nucleotide sequence variation in bovine MSTN in 410 New Zealand (NZ) Holstein-Friesian × Jersey (HF × J)-cross cows. These cows ranged from 3 to 9 years of age and over the time studied, produced an average 22.53 ± 2.18 L of milk per day, with an average milk fat content of 4.94 ± 0.17% and average milk protein content of 4.03 ± 0.10%. Analysis of a 406-bp amplicon from the intron 1 region, revealed five nucleotide sequence variants (A–E) that contained seven nucleotide substitutions. Using general linear mixed-effect model analyses the AD genotype was associated with reduced C10:0, C12:0, and C12:1 levels when compared to levels in cows with the AA genotype. These associations in NZ HF × J cross cows are novel, and they suggest that this variation in bovine MSTN could be explored for increasing the amount of milk unsaturated fatty acid and decreasing the amount of saturated fatty acid.

Meat quality and carcass characteristics of Norwegian dairy and Holstein-Friesian cattle

2007 ◽  
Vol 2007 ◽  
pp. 110-110
Author(s):  
R.M. Kirkland ◽  
D.C. Patterson ◽  
B.W. Moss ◽  
T.W.J. Keady ◽  
R.W.J. Steen
Keyword(s):  
Meat Quality ◽  
Production Systems ◽  
Carcass Characteristics ◽  
Quality Parameters ◽  
Quality Characteristics ◽  
Production Data ◽  
Dairy Traits ◽  
Holstein Friesian ◽  
Friesian Cattle ◽  
Norwegian Dairy

Any evaluation of breeds or production systems for beef must consider effects on production, carcass and meat quality characteristics. Holstein-Friesian (HF) cattle are bred for dairy traits only, while Norwegian dairy cattle (NOR) have been selected with some emphasis on beef characteristics. A comparison of production data from bulls of these two breeds has been presented previously (Kirkland et al., 2005). The objective of the present study was to evaluate specific carcass and meat quality parameters of HF and NOR bulls.

scholarly journals Dysfunction of the TP53 tumor suppressor gene in lymphoid malignancies

Blood ◽  
2012 ◽  
Vol 119 (16) ◽  
pp. 3668-3683 ◽  
Author(s):  
Zijun Y. Xu-Monette ◽  
L. Jeffrey Medeiros ◽  
Yong Li ◽  
Robert Z. Orlowski ◽  
Michael Andreeff ◽  
...  
Keyword(s):  
Tp53 Gene ◽  
Suppressor Gene ◽  
Wild Type ◽  
Lymphoid Malignancies ◽  
The Status ◽  
Tp53 Tumor Suppressor Gene ◽  
P53 Activation ◽  
Apoptotic Pathways ◽  
P53 Inactivation ◽  
Tp53 Pathway

AbstractMutations of the TP53 gene and dysregulation of the TP53 pathway are important in the pathogenesis of many human cancers, including lymphomas. Tumor suppression by p53 occurs via both transcription-dependent activities in the nucleus by which p53 regulates transcription of genes involved in cell cycle, DNA repair, apoptosis, signaling, transcription, and metabolism; and transcription-independent activities that induces apoptosis and autophagy in the cytoplasm. In lymphoid malignancies, the frequency of TP53 deletions and mutations is lower than in other types of cancer. Nonetheless, the status of TP53 is an independent prognostic factor in most lymphoma types. Dysfunction of TP53 with wild-type coding sequence can result from deregulated gene expression, stability, and activity of p53. To overcome TP53 pathway inactivation, therapeutic delivery of wild-type p53, activation of mutant p53, inhibition of MDM2-mediated degradation of p53, and activation of p53-dependent and -independent apoptotic pathways have been explored experimentally and in clinical trials. We review the mechanisms of TP53 dysfunction, recent advances implicated in lymphomagenesis, and therapeutic approaches to overcoming p53 inactivation.

NAT2 and CYP1B1 genetic polymorphisms in patients with genital endometriosis depending on tolerability of melatonin

2021 ◽  
Vol 70 (4) ◽  
pp. 35-42
Author(s):  
Tatyana E. Ivashchenko ◽  
Maria I. Yarmolinskaya ◽  
Saimat S. Tkhazaplizheva
Keyword(s):  
Genetic Polymorphism ◽  
Rflp Analysis ◽  
Acetylator Phenotype ◽  
Wild Type ◽  
Melatonin Administration ◽  
Pcr Rflp ◽  
Genes Encoding ◽  
Poor Tolerance ◽  
Nat2 Gene ◽  
Rapid Acetylator

BACKGROUND: Genital endometriosis is one of the most pressing problems of modern gynecology. Melatonin is a promising drug with a potentially curative effect on endometriosis. AIM: The aim of this study was to conduct a comparative analysis of the genetic polymorphism of some genes encoding enzymes involved in melatonin metabolism. MATERIALS AND METHODS: The genetic polymorphism in the NAT2 and CYP1B1 genes encoding enzymes involved in melatonin metabolism in patients with different tolerance to this drug was analyzed by PCR-RFLP analysis. RESULTS: In patients with genital endometriosis, the presence of a wild-type allele (N) of the NAT2 gene was associated with poor tolerance of melatonin. The NAT2 (N / N) rapid acetylator phenotype combined with the low catalytic activity of CYP1B1 (C / C) occurred more frequently in endometriosis patients having poor melatonin tolerability compared to the group of patients who tolerated the therapy well. CONCLUSIONS: For patients with genital endometriosis with the wild-type (N) allele of the NAT2 gene, melatonin administration is inappropriate due to numerous side effects during the drug use.

Myristylation is required for Tyr-527 dephosphorylation and activation of pp60c-src in mitosis

1993 ◽  
Vol 13 (3) ◽  
pp. 1464-1470
Author(s):  
S Bagrodia ◽  
S J Taylor ◽  
D Shalloway
Keyword(s):  
Kinase Activity ◽  
Tyrosine Phosphatase ◽  
Src Kinase ◽  
Indirect Evidence ◽  
Membrane Localization ◽  
Wild Type ◽  
Src Tyrosine Kinase ◽  
Amino Terminal ◽  
Membrane Bound ◽  
Type C

The chicken proto-oncoprotein c-Src is phosphorylated by p34cdc2 during mitosis concomitant with increased c-Src tyrosine kinase activity. On the basis of indirect evidence, we previously suggested that this is caused by partial dephosphorylation at Tyr-527, the phosphorylation of which suppresses c-Src kinase activity. In support of this hypothesis, we now show that treatment of cells with a protein tyrosine phosphatase inhibitor, sodium vanadate, blocks the mitotic increase in Src kinase activity. Also, we show that an amino-terminal mutation that prevents myristylation (and membrane localization) of c-Src does not interfere with the p34cdc2-mediated phosphorylations but blocks both mitotic dephosphorylation of Tyr-527 (in kinase-defective Src) and stimulation of c-Src kinase activity. Furthermore, in unsynchronized cells, the kinase activity of nonmyristylated c-Src is suppressed by 60% relative to wild-type c-Src, presumably because of increased Tyr-527 phosphorylation. Consistent with this, the Tyr-527 dephosphorylation rate measured in cell homogenates is much higher for wild-type, myristylated c-Src than for nonmyristylated c-Src. Tyr-527 phosphatase activity was primarily associated with the nonsoluble subcellular fraction. These findings suggest that the phosphatase(s) that acts on Tyr-527 is membrane bound and indicate that membrane localization of c-Src is necessary for its mitotic activation by dephosphorylation of Tyr-527.

Sign in / Sign up

Export Citation Format

Share Document