Biotechnology has enabled the modification of agricultural materials in a very precise way, thereby improving productivity and yields of economically important crops. There are a number of methods available for detecting genetically modified organisms (GMOs). In the present investigation, a qualitative PCR technique has been adopted in order to discriminate between genetically modified and non-modified food products. The qualitative PCR assay employs primers specific for genetic elements that are used to generate genetically engineered agricultural crops. Two of the most common primers used for the detection of GMOs, 35S promoter and NOS 3 terminator, have been tested over a panel of 24 food products purchased from the local market. The results indicated that, out of the 24 food products tested, three products gave positive results with the 35S promoter. The NOS 3 primers gave negative results with all tested samples. INTRODUCTIONPlants are consumed directly as a whole or are processed into many types of foods. Early farmers selected and preserved plant variants that had desirable food or agronomic attributes, such as larger fruit or uniform dormancy and maturation times for seeds. The objectives of plant breeders for the improvement of agricultural crops later focused on increasing yield, improving quality and reducing production costs by, for example, identifying traits that increase resistance to pests and diseases. Recent advances in biotechnology have managed, by directly modifying genes, to alter living organisms more quickly and more precisely than has been done by nature and humans over millennia. 1 Production of genetically modified (GM) plants proceeds by inserting selected foreign genes of interest into their genomes. The process by which the foreign DNA is constructed requires the splicing of several elements to form a functional genetic unit. Modification generally includes two regulatory elements, a promoter and a terminator, as well as a gene that confers a selectable advantage. 2 Two of the most commonly used regulatory elements are the constitutive 35S promoter from the cauliflower mosaic virus and the NOS 3 terminator isolated from nopaline synthase gene (NOS) of Agrobacterium tumefaciens. These two elements are present either together or separately in most of the transformed organisms. 3,4 The cauliflower mosaic virus (CaMV) 35S promoter is constitutively
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.
Here, we sequenced the complete mitochondrial genome of 29 Egyptian river buffaloes collected in two breeding stations of Egypt. The genome is 16,357–16,359 base pairs in length and contains the 37 genes found in a typical mammalian genome. The overall base composition is A: 33.1%, C: 26.6%, G: 13.9%, and T: 26.4%. Our analyses confirm that the mitochondrial genomes of swamp and river buffaloes are divergent (mean nucleotide distance = 2.3%), and show that Indian river buffalo haplotypes cluster into three haplogroups, named RB1, RB2, and RB3 (mean distance = 0.25–0.26%) and that the 24 Egyptian buffalo haplotypes fall into RB1 (with the Bangladeshi, Chinese and Italian buffalo haplotypes) and RB2.
Enrichment of barcode databases with mitochondrial cytochrome c oxidase subunit I (COI) barcode sequences in different animal taxa has become important for identification of animal source in food samples to prevent commercial fraud. In this study, COI barcode sequence in seventy one river buffalo samples were determined, analyzed and deposited in Genbank barcode database and barcode of life database (BOLD) to contribute for construction of public reference library for COI barcode sequence in river buffalo. Moreover COI barcode sequence was used to identify the closely related buffalo groups: river buffalo, swamp buffalo, lowland anoa and African buffalo. Results indicated the success of the COI barcode in the identification of each of the tested groups. Whereas a suggested sequence of other mitochondrial segment representing two successive transfer RNA (tRNA) genes; tRNA-Threonine (MT-TT) and tRNA-Proline (MT-TP) was failed to be used as a barcode marker for differentiation between the tested buffalo groups.
The present study was performed to evaluate the effect of three natural antioxidants on the adverse effect of methotrexate (MTX) in normal liver cells. TaqMan RT-PCR technology was used to estimate the mRNA expression levels for three genes after rats injection with a single dose of 20 mg/kg b.w MTX or the same MTX dose combined with ginger, silymarin or propolis oral administration. The doses of ginger, silymarin or propolis were similar (200 mg/kg b.w) and were daily administrated to rats for 21 days before MTX injection and four days after MTX injection. The three genes were: cytochrome P450 oxidoreductase (POR) that encodes POR enzyme, caspase 3 (CASP-3) that encodes CASP-3 enzyme and interlukin 6 (IL-6) that encodes IL-6 pro-inflammatory cytokine. Results indicate that the used MTX single dose of 20 mg/kg b.w did not significantly affect POR mRNA expression level in rat liver. Moreover the administration of ginger, silymarin or propolis with MTX did not significantly increase or decrease POR mRNA expression level. Results also reveal insignificant increase in CASP-3 mRNA expression level after MTX injection and also after administration of ginger or propolis with MTX. The administration of silymarin with MTX significantly increased the CASP-3 mRNA expression level. IL-6 mRNA expression level was insignificantly upregulated after injection with MTX and also after administration of silymarin with MTX whereas a significantly upregulation in the IL-6 mRNA expression level was reported after administration of ginger or propolis with MTX.
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