The Anti-Müllerian Hormone (AMH) is a member of the transforming growth factor beta (TGF-β) superfamily, playing a significant role in cell proliferation, differentiation and apoptosis. In females, AMH is secreted throughout their reproductive life span from ovaries, whereas in males it is secreted by gonadal cells at a very early stage of testicular development. AMH is a promising marker of ovarian reserve in women and can be used to measure the female reproductive lifespan. In the present study, we cloned and sequenced the GC rich AMH gene from Indian riverine buffalo ( Bubalus bubalis) and goat ( Capra hircus ). Obtained sequences were compared to the AMH sequences of other mammals, and corresponding amino acid sequences revealed that the caprine and bovine AMH sequences are more closely related to each other than to those of other mammals. Furthermore, we analyzed the chromosomal localization of AMH genes in mammalian species to understand potential syntenic relationship. The AMH gene is localized between the sequences for the SF3A and JSRP1 genes and maintains this precise location in relation to other nearby genes. The dN/dS ratio of AMH gene did not indicate any pressure for either positive or negative selection; thus, the physiological function of the AMH gene in the reproduction of these two ruminant species remains very vital. Similar to other mammals, the AMH gene may be an important indicator for regulating female reproductive biology function in bovine, cetacean, caprine, and camelidae.
Milk protein polymorphism was analysed to improve the protein content in milk. The present study characterises the CSN1S1 gene and the effect of allelic combinations on milk composition traits in Jamunapari goats. The allelic variants obtained from sodium dodecyl sulfate polyacrylamide gel electrophoresis and polymerase chain reaction–restriction fragment length polymorphism were confirmed by cloning and sequencing. Genetic parameters were obtained from 518 records from 48 sires and 131 dams. The A, B and F alleles were observed in the population and the protein percentage in milk was significantly ( P < 0.01) affected by allelic variants. The frequencies of A, B and F alleles were 0.456, 0.503 and 0.041, respectively. The protein content in milk was highest in the goats with AB genotype followed by AA > BB > BF > AF > FF. The goats with AB genotype had a significantly ( P < 0.01) higher protein percentage in milk than goats with BF ( t = 5.311, df = 113), AF ( t = 8.13, df = 123) and FF ( t = 9.55, df = 115) genotypes. The direct heritability for protein percentage was 0.441. Parity and season of birth had significant effects ( P < 0.05) on the solids‐not‐fat percentage and lactose concentrations. The CSN1S1 AA, AB and BB genotypes should be selected for higher protein content and to improve milk quality and processing traits in Indian goats.
The transfer of genome-modification components into farm animal cells is indispensable for the production of genome-modified and transgenic farm animals. Electroporation is a physical transfection method when appropriately used; this technique is safe, simple to use, affordable, and efficient in transfecting cells from several lineages. Electroporation efficiency depends on various physical parameters, of which cell type is considered a major factor for transfection efficiency. Primary cells are generally less susceptible to transfection than other cell types due to their finite lifespan and limited expansion capacity. Previously, we custom-designed a transfection buffer to deliver exogenous genetic components into mammalian cells. In the present study, we examined the effect of the developed buffer on transfection rates and cell viability of primary somatic cells from buffalo, cattle, goats, and sheep. To achieve the aims of this study, t primary somatic cells from skin biopsies were established and were transfected with a Venus-expression vector (pAcGFPs-Venus). We noticed that transfection rates of pAcGFPs-Venus were 22.51%, 17.56%, 22.81%, and 16.16% for buffalo, cattle, goats, and sheep cells, respectively. We also noticed that cell viability and proliferation rates were better in the case of goats, sheep, and cattle cells; also, these cells have less vacuolation than buffalo cells. In addition, we also generated MSTN (myostatin) KO (Knockout) cell clones from these cell populations, in which the efficiency of single-cell clone generation was high for goats and sheep cells. In conclusion, our lab-made transfection buffer can be efficiently used to generate genome-edited or transgenic farm animals for agriculture, biomedical, and veterinary applications.
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