المؤلفون: Karima F. Mahrous, Heba A. M. Abd El -Kader, Mohamad A. Abdelhafez, Mohamad M. Aboelenin, Esraa A. Balabel, Dalia M. Mabrouk, Osama M. EL Malky, Mohamed S. Hassanane

المصدر: Journal of Genetic Engineering and Biotechnology, Vol 20, Iss 1, Pp 1-10 (2022)

مصطلحات موضوعية: Leptin, Leptin receptor, BMP4, Gene polymorphism, Repeat breeding, Egyptian buffaloes, Biotechnology, TP248.13-248.65, Genetics, QH426-470

الوصف: Abstract Background This study aimed to explore the association between polymorphisms in three genes: leptin (LEP), leptin receptor (LEPR), and BMP4, and incidence of repeat breeding in Egyptian buffaloes. Methods DNA was extracted from 160 female buffaloes, involving 108 fertile and 52 repeat breeders. Genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Sequence analysis and alignment were performed by employing NCBI/BLAST/blastn suite, to identify SNPs among different patterns and alleles. We utilized PredictSNP software to predict the non-synonymous SNPs influences on protein function. Moreover, the conservation score of the amino acids within the target proteins was computed by ConSurf server. Results The genotyping results showed that LEP and BMP4 genes were monomorphic (CC, GG) in all tested fertile and repeat breeder buffaloes. Leptin gene sequencing showed a non-synonymous C73T SNP, replacing R to C at position 25 within the leptin polypeptide (position 4 in the mature form; R4C) which is a neutral mutation, not affecting function or structure of LEP protein. For LEPR, one synonymous SNP (T102C) and two non-synonymous SNPs (A106G and C146A), triggering V967A and G954C replacements, respectively in LEPR protein. Moreover, they are neutral mutations. Sequencing results of BMP4 showed HinfI restriction site indicate fixed GG genotype (CC genotype in the anti-sense strand) in all sequenced samples. No SNPs were observed within the amplified region. Conclusion Genotyping and sequencing results of the surveyed three genes revealed that there is no association between these genes mutations and the incidence of repeat breeding in Egyptian buffaloes.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/2090-5920Test

الوصول الحر: https://doaj.org/article/3973c8878afe4045830e3891382e9f50Test

المؤلفون: Mohamed M. Abd Elrahman, Aida I. El makawy, Mohamed S. Hassanane, Sally S. Alam, Nagwa H. A. Hassan, Medhat K. Amer

المصدر: Journal of Genetic Engineering and Biotechnology, Vol 19, Iss 1, Pp 1-15 (2021)

مصطلحات موضوعية: Male fertility, mtDNA, Mutations, Sperm motility, ROS, Lipid peroxidation, Biotechnology, TP248.13-248.65, Genetics, QH426-470

الوصف: Abstract Background Asthenozoospermia is a chief reason for male seminal pathologies with an impression of around 19% of infertile patients. Spermatozoa mitochondrial DNA variations seem to link with low sperm motility. The objective of the study was to assess the relation between mitochondrial mutations and male sterility, especially in asthenozoospermia. The patient semen samples were investigated by studying the sperm physical characters; motility, viability, and morphological parameters were then classified into normozoospermia and asthenozoospermia. In addition, the level of malondialdehyde (MDA) as a bio-indicator of lipid peroxidation, seminal fructose, and total antioxidant capacity (TAC) were estimated. For molecular analysis, DNA from the semen samples was extracted using a DNA extraction kit. ND1, ND2, and ATPase6 genes were amplified by using a specific primer. After the purification procedure, each PCR product was sequenced to identify the single nucleotide polymorphisms (SNPs) in selected genes. Results A significant negative correlation between seminal plasma malondialdehyde levels and sperm motility was detected. Meanwhile, TAC analysis revealed significantly lower activity (p ≤ 0.05) in the sample of asthenozoospermic than in normozoospermic men. As regards the seminal plasma fructose, there was no significant difference in the fructose level of normozoospermia and asthenozoospermia cases. At the molecular level, 31 diverse nucleotide substitutions were recognized in mitochondrial DNA. Only ten (10) mutations led to amino acid transformation: four have deleterious effects, four are benign, and the other two have conflicting effectiveness. Conclusions This study is the first in Egypt that is concerned with studying the relationship between the mitochondrial DNA mutations in human spermatozoa of asthenozoospermic patients and fertility. The results displayed scientific indications evidenced that there is an association between mitochondrial mutations and male infertility.

وصف الملف: electronic resource

العلاقة: https://doaj.org/toc/2090-5920Test

الوصول الحر: https://doaj.org/article/86424f7abc44455c873b9e22051b9c90Test

المؤلفون: Mohamed S. Hassanane, Amal A.M. Hassan, Fatma M. Ahmed, Esteftah M. El-Komy, Khaled M. Roushdy, Nagwa A. Hassan

المصدر: Journal of Genetic Engineering and Biotechnology, Vol 16, Iss 1, Pp 83-88 (2018)

مصطلحات موضوعية: Biotechnology, TP248.13-248.65, Genetics, QH426-470

الوصف: Egyptian chickens, representing 2 breeds and 7 strains, were genotyped using the PCR-RFLP and sequencing techniques for detection of a non-synonymous dimorphism (G/A) in exon 14 of chicken Myxovirus resistance (Mx) gene. This dimorphic position is responsible for altering Mx protein’s antiviral activity. Polymerase Chain reactions were performed using Egyptian chickens DNA and specific primer set to amplify Mx DNA fragments of 299 or 301 bp, containing the dimorphic position. Amplicons were cut with restriction enzyme Hpy81. Genotype and allele frequencies for the resistant allele A and sensitive allele G were calculated in all the tested chickens. Results of PCR-RFLP were confirmed by sequencing. The three genotypes AA, AG, GG at the target nucleotide position in Mx gene were represented in all the studied Egyptian chicken breeds and strains except Baladi strain which showed only one genotype AA. The average allele frequency of the resistant A allele in the tested birds (0.67) was higher than the sensitive G allele average frequency in the same birds (0.33). Appling PCR-RFLP technique in the breeding program can be used to select chickens carrying the A allele with high frequencies. This will help in improving poultry breeding in Egypt by producing infectious disease-resistant chickens. Keywords: Egyptian chickens, Antiviral activity, Mx gene, Genotyping, PCR-RFLP

وصف الملف: electronic resource

العلاقة: http://www.sciencedirect.com/science/article/pii/S1687157X17301087Test; https://doaj.org/toc/1687-157XTest

الوصول الحر: https://doaj.org/article/6704c2ba29744b6ab1a7337ed8f0b883Test

المؤلفون: Mohamed S. Hassanane, Aida I. El Makawy, Sahar M. Helalia, Ahmed S. Abdoon, Kamal M.A. Khalil, Tharwat A. Ghanem, Amany M. Tohamy, Xiao-Feng Sun, Wei Shen

المصدر: Journal of Genetic Engineering and Biotechnology, Vol 15, Iss 2, Pp 475-482 (2017)

مصطلحات موضوعية: Buffalo embryos, Transgenic, SMGT, EGFP, DMSO, Biotechnology, TP248.13-248.65, Genetics, QH426-470

الوصف: The present study was carried out to find the best treatments for enhancing the ration of insertion of a desired gene construct (pEGFP-N1) onto the sperm of buffalo as the first step for the production of transgenic buffalo using sperm mediated gene transfer (SMGT). The tested conditions were plasmid DNA concentration, sperm concentration, transfecting agent concentration: Dimethyle sulphoxide (DMSO) and time of transfection. The study proved that the best conditions for producing transgenic embryos were incubation sperm solution its concentration is 107/ml sperm with 3% DMSO: with 20 µg/ml from the linarized DNA, for 15 min at 4 °C are the best conditions to produce transgenic buffalo embryo using sperm mediated gene transfer.

وصف الملف: electronic resource

العلاقة: http://www.sciencedirect.com/science/article/pii/S1687157X17300318Test; https://doaj.org/toc/1687-157XTest

الوصول الحر: https://doaj.org/article/367b7c5b8c434d70a3211c9ac64efed2Test