Beneficial Lactic Acid Bacteria
K. Balamurugan, U. Prithika in Pocket Guide to Bacterial Infections, 2019
Restriction fragment length polymorphism (RFLP) is characterized by the use of restriction enzymes to digest DNA and following separation of the restriction fragments according to their length by agarose gel electrophoresis (Adzitey et al. 2013). Single digestion with AciI of rpoB gene (coding for RNA polymerase β-subunit) in Leuconostoc, Oenococcus, Pediococcus, and two or three digestions (AciI, HinfI and MseI) in Lactobacillus spp. allowed to identify LAB species commonly isolated from wine (Claisse et al. 2007). Restriction patterns of the tuf gene, encoding the elongation factor Tu and universally distributed in gram-positive bacteria, derived by enzymes AluI and HaeIII could effectively differentiate closely related Lactobacillus species (Park et al. 2012). However, sometimes strain variations could not be demonstrated by the RFLP analysis. Morphologic differences (colony shape and size) were evident between Lactobacillus kefir strains ATCC 35411 and ATCC 8007, but genotypic results failed to differentiate them (Mainville et al. 2006).
Immunology of Insulin-Dependent Diabetes Mellitus
Irun R. Cohen in Perspectives on Autoimmunity, 2020
Characterization of the class II allelic gene products associated with IDDM have relied in the past on serological techniques. The associations defined are partial, and the majority of subjects with susceptibility alleles never develop IDDM. In addition, 10% of IDDM patients do not carry the susceptibility alleles defined by serological markers. Diseases which do not appear to be related to IDDM are associated with identical susceptibility alleles. Serological definition of susceptibility alleles, therefore, may be incomplete and/or insufficient. Restriction fragment length polymorphism (RFLP) is a first step in characterizing, at the DNA level, susceptibility alleles and in verifying whether alleles which have different amino acid sequences but share identical antigenic determinants are detected as different by RFLP. This approach has been developed using a DR β probe to study HLA-DR in DR4 and DR3 heterozygous individuals. An increased frequency of a Pstl 18-kilobase (kb) fragment has been observed.176 Similar studies in DR2 healthy and diabetic individuals using a DC β cDNA probe have also allowed the definition of a 2.2-kb fragment which may be related to protection against IDDM.177
DNA TECHNIQUES FOR THE AUTHENTICATION OF CHINESE MEDICINAL MATERIALS
Kevin Chan, Henry Lee in The Way Forward for Chinese Medicine, 2001
Traditional RFLP analysis is not suitable for Chinese medicinal materials as it demands large amount of intact DNA. Therefore, PCR-RFLP, a method requiring small amount of DNA for analysis is used. In this method, a defined DNA fragment is first amplified by PCR and then digested with a restriction endonuclease to generate restriction polymorphic profiles unique to the concerned species. It is desirable that the region for PCR-RFLP analysis is flanked by sequence conserved across species so that it can be readily amplified by using "universal" primers. Two of the suitable candidates for such regions are ribosomal DNA (rDNA) and large subunit of ribulose-1,5-bisphosphate carboxylase L (rbcL) gene. The PCR-RFLP of rDNA has been carried out on Glehnia and Atractylodes (Mizukami et al. 1993; 1996; Cheng et al. 1997) and we have also successfully used this approach to discriminate Panax species from their adulterants (Figure 9.3) (Ngan et al. 1998) and?
The relationship between KRAS LCS6 polymorphism and endometrium cancer
Published in Journal of Obstetrics and Gynaecology, 2020
Feyza Nur İncesu Çintesun, Özlem Seçilmiş Kerimoğlu, Ersin Çintesun, Süleyman Nergiz, Hasan Acar, Çetin Çelik
The Restriction Fragment Length Polymorphism (RFLP) method is a method which allows the examination of DNA profiles formed by separation in the agarose gel electrophoresis system of DNA segments and fragmentation with a restriction endonuclease enzyme specific to genomic DNA. By identifying specific sequences in the DNA of these enzymes, which are found in bacteria, the fragmentation process occurs in the identified region or in another specific sequence outside this region. Genotyping of single nucleotide polymorphisms occurs in two steps with this method. The first step is based on PCR product sequencing or enzyme fragmentation after gel electrophoresis for the genotyping of the polymorphic region by hundreds of thousands of proliferations with PCR of approximately 200 base pairs around the single nucleotide polymorphism (SNP) in the DNA region (Linn and Arber 1968).
Evaluation of genetic polymorphisms at 21 autosomal STR loci in Ramgharia Sikh population of Punjab, India
Published in Annals of Human Biology, 2022
Amandeep Kaur Bhambara, Abhishek Singh, Vivek Sahajpal, Mukesh Thakur, Deepika Bhandari, Shivkant Sharma, Mukesh Kumar Thakar
India, being a diverse country with an accounted population of about 1.2 billion (Census of India, 2011), portrays a significantly colourful canvas comprising of novel assimilation of various ethnic groups differing in terms of their communities, cultures, and religions. Several researchers and philosophers have been engrossed for centuries in studying the diverse patterns regarding human appearances and forms. The genetic variation among human populations was first reported in 1919 by Ludwik and Hanka Hirszfeld, pioneers in the field of blood typing (Allan 1963). However, the variations studied in blood groups were not sufficient to identify a particular individual from a gene pool. Thus, this area of interest was revolutionised by detecting variations in the human gene pool at the DNA level (Kandpal et al. 2011). In the early days, RFLP (Restriction Fragment Length Polymorphism) analysis was employed for DNA analysis followed by the PCR-based assays e.g. SNPs, VNTRs, and Short Tandem Repeats (STRs). By the mid-1990s, forensic DNA testing introduced multiple STR markers in single multiplexed reactions (Gusmão et al. 2006).
Diagnostic markers for glaucoma: a patent and literature review (2013-2019)
Published in Expert Opinion on Therapeutic Patents, 2019
Silvia Bua, Claudiu T Supuran
Mutations in the myocilin gene called p. Q368X were detected by restriction fragment length polymorphism (RFLP) analysis. RFLP exploits variations in homologous DNA sequences, known as polymorphisms, to distinguish individuals, populations, or species, or to pinpoint the locations of genes within a sequence. In RFLP analysis, a DNA sample is digested into fragments by one or more restriction enzymes, and the resulting restriction fragments are then separated by gel electrophoresis according to their size. Undoubtedly, RFLP analysis has been an important early tool for genome mapping, localization of genes for genetic disorders, determination of risk for disease, and paternity testing. However, restriction analysis is a relatively expensive and time-consuming method, which might last over 3 days. As a result, faster and cheaper DNA sequencing technologies have been developed nowadays which made RFLP obsolete.
Related Knowledge Centers
- DNA
- DNA Profiling
- DNA Sequencing
- Gel Electrophoresis
- Homology
- Molecular Biology
- Restriction Enzyme
- Restriction Site
- Gene Polymorphism
- Gene