Showing 3 results for Pcr-Rflp
Gh. Elyasi, Dj. Shodja, M.r. Nassiry, A. Tahmasebi, O. Pirahary,
Volume 9, Issue 2 (7-2005)
Abstract
In modern programmes of animal breeding, the polymorphisms of the milk proteins can be used as marker systems. Beta-Lactoglobulin is the major milk whey protein in the ruminants. The BLG coding gene located on ovine chromosome 3. This protein, synthesis in the mammary glands during pregnancy and the lactation stages. Studies have indicated that this protein is polymorphic in the many breeds of sheep. This is the result of a single base pair substitution in the Beta-Lactoglobulin gene that also rises to the RsaI restriction fragment length polymorphism (RFLP). The aim of this work was to analyze the genotype distribution of Beta-Lactoglobulin in sheep. Blood samples were supplied from 142 sheep of the 5 breeds (Ghezel, Afshari, Moghani, Makoii and Arkharmerino). Genomic DNA was extracted from the 200ul blood sample according to Boom et al. (1989) method modified by Shaikhayev (1995). The Gel monitoring and the spectrophotometeric methods were used for determination of the DNA quality and quantity. the Primers BLG5 and BLG3 amplified a 452 bp fragment from the exon II of the ovine Beta-Lactoglobulin gene. the Products of the amplification were recognized by the electrophoresis on the 1% agarose gel stained with ethidium bromide. The RsaI enzyme was used for restriction of the PCR products. The digested products were separated by the electrophoresis on the 8% nondenaturant polyacrylamide gel and visualized after staining with the ethidium bromide on UV transillumination. The popGen32 software (ver.1.31) was used to estimate the allele and the genotype frequencies, the Hardy-Weinberg equilibrium and dendrogram of the genetic distance. The frequency of A-allele in Ghezel, Afshari, Moghani, Makoii and Arkharmerino breeds was 56%, 34%, 36%, 53% and 48% respectively. The populations were in Hardy-Weinberg equilibrium except to Afshari breed. The lowest genetic distance was observed between Moghani and Afshari breeds and the highest genetic distance between Ghezel and Afshari breeds. The results of this study indicated that PCR-RFLP is an appropriate tool for evaluating genetic variability in sheep.
E. Mahdikhani Moghadam, A. Kheiri, M. Mohammadi,
Volume 10, Issue 4 (1-2007)
Abstract
To carry out this study, total DNA was extracted from eggs and from second stage juveniles of several populations of Meloidogyne javanica and Meloidogyne incognita, using phenol / chloroform method. Following extraction, DNA was electrophoresed on 1% agarose gel to determine its quality and quantity. A specific primer pair (C2F3 / 1108 23 and 20 nucleotides, respectively) was used to discriminate M. javanica from M. incognita populations using polymerase chain reaction (PCR). Primer annealing sites were located in the 3′ portion of mitochondrial gene encoding cytochrome oxidase subunit II (COII) and in the 16S rRNA gene. Following PCR amplification, electrophoresis of amplified DNA showed 1.7 kb fragment in populations of both species. Digestion of 1.7 kb amplified product with HinfI restriction endonuclease resulted in the generation of two DNA fragments of 0.7 and 1.0 kb in M. javanica and three DNA fragments of 0.3, 0.4 and 1.0 kb in M. incognita. There were no differences in the digestion patterns among various populations of each species examined.
M Talebi, M Bahar, Gh Saeedi, A Mohamadi,
Volume 13, Issue 47 (4-2009)
Abstract
To characterize the geographical distribution of medicago-nodulating rhizobia in western regions of Iran, 950 Sinorhizobium isolates were trapped from a combination of two local alfalfa populations (Hamedani, Nikshahri) together with a foreign cultivar ( Kodi) and soil samples from eight sites across Kurdestan, Kermanshah, Eastern Azarbayjan and Lorestan provinces. Also, a total of 45 isolates were obtained from nodules of naturally grown Melilotus officinalis (14 isolates) and Trigonella foenum-graecum (31 isolates) plants in Isfahan. On the basis of PCR partial amplification of the plasmid born nod box gene and chromosomal mucR gene of the isolates,16S ribosomal DNA PCR-restriction fragment length polymorphism analysis, and the nucleotide sequence, three isolates from alfalfa, seven isolates from M. officinalis and 13 isolates from T. foenum-graecum were proved to be Sinorhizobium medicae. The remaining isolates (943 from alfalfa, seven from M. officinalis and 18 from T. foenum-graecum) were identified as S. melilloti. Both species, S. meliloti and S. medicae, were recovered from nodules of all the hosts although S. meloti was clearly more dominant in nodulating different populations of alfalfa. Taken together, these results indicated that the abundance of S. meliloti is independent of the site of isolation and have a wide geographical distribution. In this study, the banding pattern resulting from PCR amplification of 16S rRNA gene, followed by digestion with Rsa I, clearly differentiated S. meliloti and S. medica strains, showing that PCR-RFLP is an appropriate method to discriminate medicago-nodulating rhizobian with relative rapidity.
