An easy and accurate discrimination method of Ryukyu wild boar (Sus scrofa riukiuanus) by PCR-RFLP analysis using Hinf I site of D-loop region of mitochondorial DNA

Abstract Itoi, S., Saito, T., Shimojo, M., Washio, S., and Sugita, H. 2007. Identification of Girella punctata and G. leonina by PCR-RFLP analysis. – ICES Journal of Marine Science, 64: 328–331. Two Girella species, Girella punctata and G. leonina, are sympatric sister species with an extensive overlap in their respective distributions on shallow rocky reefs from Hong Kong to the south of the Japanese Islands. Juveniles of the two species cannot be discriminated easily on the basis of external characters. In this study, after morphological identification of the species, sequencing analysis was carried out for the partial 16S ribosomal RNA gene and for the D-loop region in mitochondrial DNA. A total of 109 specimens was examined. Restriction site mapping of the sequences suggested that the electrophoretic patterns of polymerase chain reaction (PCR)-based restriction fragment length polymorphism (RFLP) analysis of the gene products would produce a species-specific banding pattern. Subsequently, the PCR-RFLP analysis showed that the method was as effective for separating the two morphologically similar species of the genus Girella as the sequencing analysis.

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Polymorphisms of genes required for methionine synthesis and DNA methylation influence mitochondrial DNA methylation

Epigenomics ◽

10.2217/epi-2020-0041 ◽

2020 ◽

Vol 12 (12) ◽

pp. 1003-1012

Author(s):

Andrea Stoccoro ◽

Pierpaola Tannorella ◽

Lucia Migliore ◽

Fabio Coppedè

Keyword(s):

Dna Methylation ◽

Mitochondrial Dna ◽

Exploratory Study ◽

High Resolution Melting ◽

Late Onset ◽

Loop Region ◽

Pcr Rflp ◽

D Loop ◽

One Carbon Metabolism ◽

Methionine Synthesis

Aim: Impaired methylation of the mitochondrial DNA and particularly in the regulatory displacement loop (D-loop) region, is increasingly observed in patients with neurodegenerative disorders. The present study aims to investigate if common polymorphisms of genes required for one-carbon metabolism ( MTHFR, MTRR, MTR and RFC-1) and DNA methylation reactions ( DNMT1, DNMT3A and DNMT3B) influence D-loop methylation levels. Materials & methods: D-loop methylation data were available from 133 late-onset Alzheimer’s disease patients and 130 matched controls. Genotyping was performed with PCR-RFLP or high resolution melting techniques. Results: Both MTRR 66A > G and DNMT3A -448A > G polymorphisms were significantly associated with D-loop methylation levels. Conclusion: This exploratory study suggests that MTRR and DNMT3A polymorphisms influence mitochondrial DNA methylation; further research is required to better address this issue.

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PCR-based genetic identification of marlin and other billfish

Marine and Freshwater Research ◽

10.1071/mf98007 ◽

1998 ◽

Vol 49 (5) ◽

pp. 383 ◽

Cited By ~ 8

Author(s):

B. H. Innes ◽

P. M. Grewe ◽

R. D. Ward

Keyword(s):

Mitochondrial Dna ◽

Genetic Test ◽

Rflp Analysis ◽

Restriction Enzymes ◽

Genetic Identification ◽

Dna Molecule ◽

Pcr Rflp ◽

D Loop ◽

Specific Variation ◽

Species Specific

A genetic test was developed for the identification of the six species of billfish found in Australian waters (black marlin, Indo–Pacific blue marlin, striped marlin, Indo–Pacific sailfish, shortbill spearfish and broadbill swordfish). The test was based on the PCR–RFLP analysis of a 1400 bp region of the mitochondrial DNA molecule, the d-loop, using four restriction enzymes (Hinf I, Rsa I and Sau3A I andTaq I). A total of 33 composite haplotypes were observed among 160 fish; all were species-specific. Three of the species—black marlin, striped marlin and broadbill swordfish—showed sufficient intra-specific variation to be useful in population structure analyses.

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