Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 7 de 7
Filtrar
Más filtros










Base de datos
Intervalo de año de publicación
1.
Anim Biotechnol ; 31(2): 135-141, 2020 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-30905313

RESUMEN

In the present study, the potential of different grades of cumulus-oocyte complexes (COCs) for in vitro maturation (IVM) and embryonic development was assessed. Further, the association of the expression pattern of anti-apoptotic Mcl-1 and pro-apoptotic Bax genes in embryonic development was analyzed. Abattoir derived oocytes were graded into grade A and B based on surrounding cumulus rings. Out of 1050 ovaries, a total number of 770 and 1360, were of grade A and B COCs, respectively, were aspirated. After IVM, grade A COCs had a significantly higher number of polar bodies (92.04 ± 0.60%) as compared to grade B (85.88 ± 0.46%). On IVF and embryo culture, grade A COCs produced the significantly higher rate of cleavage and blastocyst (90.44 ± 0.71% and 41.55 ± 0.96%) as compared to grade B COCs (79.77 ± 0.76% and 30.44 ± 0.96%). The transcriptional analysis of apoptotic genes expression by Real-time PCR revealed a significantly higher expression of Mcl-1 gene in embryos of grade A as compared to grade B, whereas, the relative expression of Bax gene was down-regulated in grade A than grade B embryos. Thus it was concluded that the pattern of apoptotic genes expression in early-stage embryos can be used as a marker gene to predict the developmental competence of COCs.


Asunto(s)
Búfalos/embriología , Células del Cúmulo/fisiología , Desarrollo Embrionario/fisiología , Regulación del Desarrollo de la Expresión Génica/fisiología , Técnicas de Maduración In Vitro de los Oocitos/veterinaria , Oocitos/fisiología , Animales , Apoptosis/fisiología , Fertilización In Vitro/veterinaria , Técnicas de Maduración In Vitro de los Oocitos/métodos , Proteína X Asociada a bcl-2/genética , Proteína X Asociada a bcl-2/metabolismo
2.
Gene Expr Patterns ; 25-26: 118-123, 2017 11.
Artículo en Inglés | MEDLINE | ID: mdl-28669682

RESUMEN

The success of Somatic cell nuclear transfer (SCNT) primarily depends on the extent of reprogramming of donor cells genome. The error of reprogramming may lead to inappropriate expression of embryonic genes at any stage of development. Under the present study the relative expression of different genes related to pluripotency (Oct-4 and Nanog), growth factors (IGF-2 and IGF-2R) and DNA methyltransferase gene (Dnmt-1) was evaluated in SCNT embryos at 8-16 cells, morula and blastocyst stages as compared to IVF group. In SCNT, significantly higher degree of relative expression was observed for Oct-4 in morula (1.41) and blastocysts (1.14) as compared to 8-16 cells (referral stage) whereas in IVF, a lower expression was observed at morula (0.82) stage. The expression of Nanog in SCNT embryos was increased significantly in morula (2.23) and decreased subsequently in blastocyst (0.56), whereas it was increased significantly from 8 to 16 cells to morula (1.62) and blastocyst (4.5) of IVF group. The IGF-2 and IGF-2R showed significantly higher expression rates in morula and blastocysts of SCNT (6.56, 5.90 and 1.11, 1.4) and IVF (8.69, 8.25 and 2.96, 3.91) embryos, respectively as compared to referral stage. The expression of Dnmt-1 was significantly higher in SCNT morula (1.29) and blastocyst (1.15) however in IVF, it was similar in 8-16 cells stage and morula but, higher in blastocyst (1.58). The dissimilar pattern of gene expression of SCNT might be a consequence of incomplete reprogramming of donor nucleus which resulted into lower blastocyst rate of SCNT as compared to IVF embryos.


Asunto(s)
Clonación de Organismos/veterinaria , Perfilación de la Expresión Génica/veterinaria , Genes del Desarrollo , Cabras/embriología , Técnicas de Transferencia Nuclear/veterinaria , Animales , Técnicas de Reprogramación Celular , Clonación de Organismos/métodos , ADN (Citosina-5-)-Metiltransferasa 1/genética , Regulación del Desarrollo de la Expresión Génica , Cabras/genética , Factor II del Crecimiento Similar a la Insulina/genética , Proteína Homeótica Nanog/genética , Factor 3 de Transcripción de Unión a Octámeros/genética , Receptor IGF Tipo 2/genética
3.
Mitochondrial DNA B Resour ; 1(1): 949-950, 2017 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-33490429

RESUMEN

Domestic pig (Sus scrofa domesticus) is one of the important farm animal contributing 7% of the country's total animal protein sources in India. In the present study, random hexamer primer was used to amplify the complete mitochondrial genome of Central Indian domestic pig and resolved the complete mitochondrial sequence by shotgun sequencing followed by de novo assembly in MIRA version 4.0.5. The sequence assembly revealed to be 15,827 bp mitogenome of pig (accession no. KT965278). The mitogenome in the present study has 99% homology with previously reported mitogenome of pigs from different parts of the world. The present study is the first report of complete sequence of mitogenome of pig from Indian subcontinent. Mitogenome analysis by MITOS web server revealed similarity of gene order, organization with the other pig breeds and vertebrates, comprising of 13 protein-coding genes, 22 tRNAs, 2 rRNAs and a control region. It was concluded that modified random hexamer can be successfully used for whole mitogenome sequencing using NGS without designing mitogenome-specific primer, thereby reducing cost and labor.

4.
Anim Biotechnol ; 27(1): 44-51, 2016.
Artículo en Inglés | MEDLINE | ID: mdl-26690650

RESUMEN

Knockdown of myostatin gene (MSTN), transforming growth factor-ß superfamily, and a negative regulator of the skeletal muscle growth, by RNA interference (RNAi), has been reported to increase muscle mass in mammals. The current study was aimed to cotransfect two anti-MSTN short hairpin RNA (shRNA) constructs in caprine fetal fibroblast cells for transient silencing of MSTN gene. In the present investigation, approximately 89% MSTN silencing was achieved in transiently transfected caprine fetal fibroblast cells by cotransfection of two best out of four anti-MSTN shRNA constructs. Simultaneously, we also monitored the induction of IFN responsive genes (IFN), pro-apoptotic gene (caspase3) and anti-apoptotic gene (MCL-1) due to cotransfection of different anti-MSTN shRNA constructs. We observed induction of 0.66-19.12, 1.04-4.14, 0.50-3.43, and 0.42-1.98 for folds IFN-ß, OAS1, caspase3, and MCL-1 genes, respectively (p < 0.05). This RNAi based cotransfection method could provide an alternative strategy of gene knockout and develop stable caprine fetal fibroblast cells. Furthermore, these stable cells can be used as a cell donor for the development of transgenic cloned embryos by somatic cell nuclear transfer (SCNT) technique.


Asunto(s)
Fibroblastos/metabolismo , Técnicas de Silenciamiento del Gen/métodos , Miostatina/genética , ARN Interferente Pequeño/genética , Transfección/métodos , Animales , Apoptosis , Proteínas Reguladoras de la Apoptosis/genética , Caspasa 3/genética , Células Cultivadas , Feto/citología , Fibroblastos/citología , Cabras , Interferones/metabolismo , Miostatina/metabolismo , ARN Interferente Pequeño/metabolismo
5.
Appl Biochem Biotechnol ; 177(2): 486-97, 2015 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-26234434

RESUMEN

Skeletal muscle is the major component of lean tissue that is used for consumption, and myostatin is a negative regulator of skeletal muscle growth. Downregulation of this gene therefore offers a strategy for developing superior animals with enhanced muscle growth. Knockdown of myostatin was achieved by RNA interference technology. The anti-myostatin shRNA were designed and stably transfected in caprine fibroblast cells. The reduced expression of target gene was achieved and measured in clonal fibroblast cells by real-time PCR. Two single-cell clones induced significant decrease of myostatin gene expression by 73.96 and 72.66 %, respectively (P < 0.05). To ensure the appropriate growth of transfected cell, seven media were tested. The best suited media was used for transfected fibroblast cell proliferation. The findings suggest that shRNA provides a novel potential tool for gene knockdown and these stably transfected cells can be used as the donor cells for animal cloning.


Asunto(s)
Fibroblastos/metabolismo , Cabras/genética , Miostatina/genética , ARN Interferente Pequeño/metabolismo , Transfección , Animales , Secuencia de Bases , Recuento de Células , Proliferación Celular , Supervivencia Celular , Fibroblastos/citología , Regulación de la Expresión Génica , Silenciador del Gen , Datos de Secuencia Molecular , Miostatina/metabolismo , ARN Interferente Pequeño/química , ARN Interferente Pequeño/genética , Factores de Tiempo
6.
Anim Biotechnol ; 23(3): 174-83, 2012.
Artículo en Inglés | MEDLINE | ID: mdl-22870872

RESUMEN

RNAi is an evolutionary conserved, highly efficient, and cost effective technique of gene silencing. It holds considerable promise and success has been achieved both in vitro and in vivo experiments. However, it is not devoid of undesirable side effects as dsRNA can trigger the immune response and can also cause non-specific off-target gene silencing. In the present study, silencing of myostatin gene, a negative regulator of myogenesis, was evaluated in caprine fetal fibroblasts using three different shRNA constructs. Out of these three constructs, two constructs sh1 and sh2 showed, 72% and 50% reduction (p<0.05) of myostatin mRNA, respectively. Efficient suppression (42-86%) of MSTN gene (p<0.05) was achieved even by reducing the concentration of shRNA constructs. The induction of classical interferon stimulated gene (Oligoandenylate Synthetase-1, OAS-1) was studied to analyze the immune response against shRNAs. Notably, a reduction in the potency of shRNAs to induce interferon response was observed at lower concentration for OAS1 gene. The results obtained in the study would be helpful in the abrogation of the bystander effects of RNAi for long term stable expression of anti-MSTN expression constructs in the muscle.


Asunto(s)
Cabras/genética , Cabras/inmunología , Interferones/biosíntesis , Miostatina/antagonistas & inhibidores , Miostatina/genética , 2',5'-Oligoadenilato Sintetasa/genética , Animales , Secuencia de Bases , Biotecnología , Células Cultivadas , Cartilla de ADN/genética , Feto/inmunología , Fibroblastos/inmunología , Técnicas de Silenciamiento del Gen , Cabras/crecimiento & desarrollo , Desarrollo de Músculos/genética , Desarrollo de Músculos/inmunología , Interferencia de ARN , ARN Interferente Pequeño/genética , Reacción en Cadena en Tiempo Real de la Polimerasa
7.
Acta Vet Hung ; 60(1): 165-74, 2012 Mar.
Artículo en Inglés | MEDLINE | ID: mdl-22366141

RESUMEN

In order to detect infectious bursal disease virus (IBDV), bursal tissue was collected from 10 IBD-suspected birds from a 30-day-old, IBDV-vaccinated commercial broiler chicken flock of 2000 birds exhibiting clinical signs suggestive of infectious bursal disease (IBD). The presence of IBDV was confirmed by partial amplification of the VP2 gene by reverse transcription and polymerase chain reaction. Isolates were identified as very virulent strains of IBDV (vvIBDV) by nucleotide sequence analysis. The comparison of the VP2 nucleotide sequences among the isolates revealed the presence of single-nucleotide polymorphisms in the VP2 gene of IBDV in the same flock. The comparative analysis indicated that these viruses were genetically close to the vvIBDVs previously detected in India. Our analysis provided information about the existence of vvIBDV in Central India.


Asunto(s)
Infecciones por Birnaviridae/veterinaria , Pollos , Brotes de Enfermedades/veterinaria , Virus de la Enfermedad Infecciosa de la Bolsa/patogenicidad , Enfermedades de las Aves de Corral/virología , Animales , Secuencia de Bases , Infecciones por Birnaviridae/epidemiología , Infecciones por Birnaviridae/virología , ADN Complementario/genética , India/epidemiología , Virus de la Enfermedad Infecciosa de la Bolsa/genética , Datos de Secuencia Molecular , Enfermedades de las Aves de Corral/epidemiología , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/veterinaria , Proteínas Estructurales Virales/genética , Virulencia
SELECCIÓN DE REFERENCIAS
DETALLE DE LA BÚSQUEDA
...