Restriction Fragment Length Polymorphism Linkage Map for Arabidopsis thaliana

We have constructed a restriction fragment length polymorphism linkage map for the nuclear genome of the flowering plant Arabidopsis thaliana. The map, containing 90 randomly distributed molecular markers, is physically very dense; >50% of the genome is within 1.9 centimorgans, or approx 270 kilobase pairs, of the mapped DNA fragments. The map was based on the meiotic segregation of markers in two different crosses. The restriction fragment length polymorphism linkage groups were integrated with the five classically mapped linkage groups by virtue of mapped mutations included in these crosses. Markers consist of both cloned Arabidopsis genes and random low-copy-number genomic DNA clones that are able to detect polymorphisms with the restriction enzymes EcoRI, Bgl II, and/or Xba I. These cloned markers can serve as starting points for chromosome walking, allowing for the isolation of Arabidopsis genes of known map location. The restriction fragment length polymorphism map also can associate clones of unknown gene function with mutant phenotypes, and vice versa

Analysis of bovine growth hormone gene polymorphism of local and Holstein cattle breeds in Kerman province of Iran using polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP)

Bovine growth hormone (bGH) gene is a part of the multiple gene family that contains prolactin and placental lactogens. Also, variations in introns have potential usefulness as genetic markers and could help in the genetic improvement of populations. Genomic DNA was isolated from blood samples of two local herds (53 animals) and two Holstein herds (50 animals). Genomic DNA samples were genotyped for the GHI-AluI polymorphism by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP). A 211 bp (bGH) gene exon 5 segment was amplified by PCR using bovine specific primers. RFLPs in this segment were studied using AluI restriction enzyme. The frequencies of V and L alleles in the local and Holstein herds were 0.2 and 0.65, respectively. For both herds, significant difference from the Hardy-Weinberg equilibrium was observed.Key words: Growth hormone, polymorphism, polymerase chain reaction restriction fragment length polymorphism, local herds, Holstein herds

Species Differentiation Of Fish Samples By Restriction Fragment Length Polymorphism Analysis Of Cytochrome B Gene

Metode pengukuran polimorfisme fragmen hasil pemotongan produkreaksi polimorfik berantai oleh enzim restriksi spesifik (polymerase chainreaction-restriction fragment length polymorphism, RFLP-PCR) telah digunakanuntuk membedakan beberapa jenis ikan mentah. Situs cytochrome b mitokondria,yang diamplifikasi oleh primer universal, dipotong menggunakan empat enzimrestriksi (Bfa I, Hinf I, Msp I, Mbo II) sehingga dapat dianalisa fragment-fragmentpendeknya. Hasil yang diperolah dari pemotongan oleh enzim restriksi tersebutternyata dapat digunakan untuk membedakan tiap jenis ikan sampel. Hasilpenelitian ini menunjukkan bahwa PCR dan RFLP-PCR merupakan metode yangsensitif dan dapat dilakukan dalam waktu singkat untuk membedakan berbagaijenis ikan mentah

Role of Molecular Markers and Importance of SNP for the Development of Cotton Programs

Cotton is an important commercial cash crop and cultivated worldwide. It is very important for improvement of desirable traits for development of cotton crop. In this review paper we have discussed the overall Molecular markers and advance techniques with their utilization in cotton improvement programmers. Molecular markers have reliable results and performance increased research of cotton breeding programs. Molecular markers are used to analyze genomic variations, association mapping, fingerprinting and genetic diversity in cotton crop.SNP markers have many advantages for genotyping of large populations as compared to previous marker systems. It is more advance and efficient processing technique. With the help of SNP technique we get more accurate results as compared to other markers in a short time. Overall DNA markers are used in cotton include Restriction Fragment Length Polymorphism (RFLP), Random Amplified Polymorphic DNA (RAPD), Amplified Fragment Length Polymorphism (AFLP), Simple Sequence Repeats (SSR) and Single Nucleotide Polymorphism, with their history current development, implication and importance in cotton breeding. Keywords PCR Polymerase chain reaction QTLs Quantitative trait loci RAPD Random amplified polymorphic DNA RFLP Restriction fragment length polymorphism RIL Recombinant inbred line SNPs Single nucleotide polymorphisms SSR Simple sequence repea

Association of Restriction Fragment Length Polymorphisms among Maize Inbreds with Agronomic Performance of Their Crosses

Restriction fragment length polymorphisms (RFLPs) have been suggested as molecular markers to facilitate improvement of agronomic traits in maize (Zea mays L.). The objective of this study was to evaluate the utility of RFLP data in elucidating heterotic patterns among maize lines. Eight maize inbred lines and their 28 singlecross hybrids werevaluated for grain yield at two Iowa locations in each of 2 yr in a randomized-complete block design. The diallel mating design permitted estimation of general and specific combining ability effects. Restriction fragment length polymorphism analysis of inbred lines included five restriction enzymes and five eDNA and 28 genomic clones distributed over the maize genome. Restriction fragment length polymorphism patterns of crosses were predicted from analysis of the inbred parents. Genetic distances between inbred lines were estimated as modified Rogers\u27 distance (MRD). Grain yield and specific combining ability were significantly correlated with MRD for six of the 10 chromosomes. Dispersion of inbred lines and hybrids for RFLP allele frequencies was generally consistent with expectations based on known pedigrees. Results from this study suggest RFLP analysis as a potential alternative to field testing when attempting to assign maize inbred lines to heterotic groups

Analysis of two single-nucleotide polymorphisms (SNPs) located in exon 1 of kappa-casein gene (CSN3) in Martina Franca donkey breed

The aim of this study is to assess genetic polymorphism at two loci in the exon 1 of the kappa-casein gene (CSN3) in Martina Franca donkey breed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis. Martina Franca donkey was derived from the Catalan donkey brought to Apulia at the time of the Spanish rule. This donkey is tall and well built and has good temperament. Both considered loci were found to be monomorphic in the considered population. At CSN3/PstI locus, all the animals were genotyped as AA since no AG and GG animals were found in the population. A similar result was found at CSN3/BseYI locus: all the donkeys were monomorphic and genotyped as AA. As a consequence, only one out of nine possible combined genotype (AAAA) was detected.Key words: Martina Franca donkey, kappa-casein gene (CSN3), gene polymorphism, polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)

Kappa-casein gene polymorphism in Holstein and Iranian native cattle by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP)

Caseins amount to nearly 80% of the protein output in cow milk. Caseins are biologically important proteins and they are also a raw material for the cheese making industry. The aim of this study was to identify kappa-casein genotype in Holstein and Iranian native cattle. DNA was extracted from 457 blood samples of 247 Holstein and 210 native cattle for identification and genotyping of kappa-casein gene by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) assay using HindIII and TaqI restriction endonucleases. The PCR product of the specific primer K-F and K-R gives the 379 bp specific band. Digestion of 379 bp fragment by restriction endonuclease HindIII generated two fragments of 156 and 223 bp. Result of the cut with this enzyme indicate there genotypes AA, AB and BB in the samples. Also, the amplified DNA (379 bp) from the samples remained undigested by TaqI restriction enzyme. These findings suggest that BB genotype could be a good factor for increase of fat and protein content of milk.Key words: Polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP), kappa-casein gene, genotyping, Holstein, native cattle

Molecular markers for genetic diversity and phylogeny research of Brazilian sheep breeds

Brazilian sheep descended from several breeds brought to the New World by Portuguese and Spanish colonists, and they have evolved and adapted to local climatic variations and acquired tolerance or resistance to many diseases. Molecular markers are widely used in analyzing genetic variability, and markers such as amplified fragment length polymorphism (AFLP), restriction fragment length polymorphism (RFLP), microsatellite, mtDNA and single nucleotide polymorphism (SNP) have facilitated the characterization of genetic diversity and population structure, and in the investigation of the natural history, behavior, and evolution of several sheep breeds. In this context, we present here a review of the uses of molecular markers in ecological and conservation research of Brazilian sheep breeds.Key words: DNA polymorphism, genetic conservation, Ovis aries

A restriction fragment length polymorphism map of Cochliobolus heterostrophus

An RFLP map was constructed for Cochliobolus heterostrophus, the causal agent of southern corn leaf blight, to facilitate the cloning of pathogenicity genes. A total of 128 RFLP and 4 phenotypic markers were analyzed. Of these, 126 were linked to other markers and delimited 944 cM, or, at a maximum, 58% of the total map length. Several RFLPs were found tightly linked to Tox1, a pathogenicity gene. A previously hypothesized translocation tightly linked to Tox1 was also detected;The numbers and sizes of the chromosomes of the two strains crossed to make the RFLP map were investigated using pulsed field gel electrophoresis. Based on our best estimate, 15 A chromosomes and 1 B chromosome were in B30.A3.R.45 whereas 15 A chromosomes were in Hm540. The size range of the chromosomes was from about 1.3 Mb to about 3.7 Mb. The physical genome size was estimated to be 35-40 Mb. The average kb/cM ratio was calculated to be roughly 22-25, smaller than that of most organisms. The low kb/cM value should make the RFLP map a very powerful tool for cloning genes using various strategies;Both probes for making the RFLP map and cloned genes from C. heterostrophus were hybridized to the separated chromosomes to associate the linkage groups and cloned genes with individual chromosomes. The reciprocal translocation associated with Tox1 and other differences in chromosome arrangement between the parents were also verified by this method

Association between A59V polymorphism in exon 3 of leptin gene and reproduction traits in cows of Iranian Holstein

We used the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique to screen for DNA polymorphisms of the leptin gene in 255 cows of Iranian Holstein. Amplified region is located in exon 3 of leptin gene. The genomic bovine leptin sequences, which consist of three exons, were obtained from GeneBank (Accession number U50365). Genotype frequencies in all herds were 0.588, 0.388 and 0.024 for AA, AB and BB, respectively, and allelic frequencies were 0.782 and 0.218 for A and B, respectively. We investigated effect of A59V polymorphism in the leptin gene on three reproduction traits. Significances of the genotype effects were tested using approximated F-statistic provided by SAS (v.8, GLM procedure). This study showed that genotype had no effect on open days and calving interval (NS) but had significant effect on length of pregnancy (P < 0.01). Animals with the AA genotype had higher length of pregnancy than other genotypes.Keywords: Leptin, Iranina Holstein, polymerase chain reaction-restriction fragment length polymorphism, reproduction traitAfrican Journal of Biotechnology Vol. 9(36), pp. 5997-6000, 6 September, 201