Screening of Three Different Alleles of mtDNA (G709A, G3496T, A3537G) in Subpopulation of UKM Students
DOI:
https://doi.org/10.18034/mjmbr.v5i1.447Keywords:
mtDNA polymorphisms, mutation, Allele G709A, Allele 3496T, Allele A3537GAbstract
Human DNA consists of nucleus DNA (nDNA) and mitochondrial DNA (mtDNA). Both are valuable in medicine and forensic genetics but in this project, single nucleotide polymorphisms (SNPs) in mtDNA are used to trace the mutation occurred. Mutations in the sequence of alleles can lead to haplogroup variation and also certain diseases. The purpose of this study is to screen of mutations on alleles G709A, G3496T, and A3537G in Malay population of The National University of Malaysia (UKM) students. These SNPs lie in the ND1 (nitrogen dehydrogenase subunit 1) coding region, and the reports state that these three alleles are prone to mutate. From MitoMap Web site, the mutations of these alleles are reported to have potential in causing several diseases with the collaboration of other SNPs mutation. Allele G709A is reported to have an association with hearing loss and Leber Hereditary Optic Neuropathy (LHON) while allele G3496T is associated to LHON only. Allele A3537G is related to diabetes. A total of 100 DNA samples were collected from Malay students of UKM and preserved on FTA card to be purified later. The concentration of the DNA on the purified FTA card was between 10μM to 20μM. An attempt was made by amplifying those three loci from the genomic DNA. The amplified product was detected and separated using 1% gel electrophoresis. Before sequencing, the PCR products were visualized under UV light using gel documentation system. All PCR products were sequenced to detect the mutation on every single position chosen. From the alignment of sequencing results, allele G709A and allele G3496T showed no mutation. Meanwhile four samples from alleles A3537G has the mutation. From the results obtained, it seems that mutations are rare in all selected alleles. It is recommended to increase the sample size and alleles selected in the future to increase the strength of the study. This study also should be applied to other populations in Malaysia such as Chinese and Indian.
Downloads
References
Bandelt H.J., Van Oven M and Salas A. 2012. Haplogrouping Mitochondrial DNA Sequences in Legal Medicine/ Forensic Genetics. International Journal of Legal Medicine 126(6): 901-916
Fu Q., Mittnik A., Johnson P.L., Bos K., Lari M., Bollongino R., Sun C., Giemsch L., Schmitz R. and Burger J. 2013. A Revised Timescale for Human Evolution Based on Ancient Mitochondrial genomes. Current Biology
Ghochani M., Nulton J., Salamon P., Frey T., Rabinovitch A and Baljon A. 2010. Tensile Forces and Shape Entropy Explain Observed Crista Structure in Mitochondria. Biophysical Journal 99(10): 3244-3254
Guo H., Zhuang X.Y., Zhang A.M., Zhang W, Yuan Y., Guo L., Yu D.D., Liu J., Yang D.K. and Yao Y.G. 2012. Presence of Mutation M. 1448t> C in a Chinese Family with Maternally inherited Essential Hypertension but No Expression of LHON. Biochimica et Biophysica Acta (BBA)-Molecular Basis of Disease
Kurelac I., Lang M., Zuntini R., Calabrese C., Simone D., Vicario S., Santamaria M., Attimonelli M., Romeo G. and Gasparre G. 2012. Searching for a Needle in the Haystack: Comparing Six Methods to Evaluate Heteroplasmy in Difficult Sequence Context. Biotechnology Advances 30(1): 363-371
Levine B. and Elazar Z. 2011. Inheriting Maternal mtDNA. Science (New York, NY) 334(6059): 1069
Pajnic I.Z., Pogorelc B.G and Balazic J. 2010. Molecular Genetic Identification of Skeletal Remains from the Second World War Kofin I Mass Grave in Slovenia. International Journal of Legal Medicine 124(4): 307-317
Prieto L., Zimmermann B., Goios A., Paneto G., Alves C., Alonso A., Fridman C., Cardoso S. and Lima G. 2011. The Ghep-Empop Collaboration on mtDNA Population Data-A New Resource for Forensic Casework. Forensic Science International: Genetics 5(2): 146-151
Rensvold J.W., Ong S.E., Jeevanathan A., Carr S.A., Mootha V.K and Pagliarini D.J. 2013. Complementary RNA and Protein Profiling Identifies Iron as a Key Regulator of Mitochondrial Biogenesis. Cell Reports 3(1): 237-245
Ruijter J., Ramakers C., Hoogars W., Karlen Y., Bakker O., Van Den Hoff M. and Moorman A. 2009. Amplification Efficiency: Linking Baseline and Bias in the Analysis of Quantitative PCR Data. Nucleic Acids Research 37(6): e45
Rydzanicz M., Wrobel M., Dominica F., Wojciech G., Witold S. and Krzysztof S. 2009. Screening of the General Polish Population for Deafness-associated Mutations in Mitochondrial 12S rRNA and tRNASer(UCN) Genes. Genetic Testing and Molecular Biomarkers 13(2): 167-172
Stalder N., Yarol N., Tozzi P., Rotman S., Morris M., Fellmann F., Schwitter J. and Hullin R. 2012. Mitochondrial A3243G Mutation with Manifestation of Acute Dilated Cardiomyopathy. Circulation Heart Failure 5(1): e1-e3
Suffiza N.A., Azuana R., Farida I. and Thomas P. 2013. Medication Adherence in Patients, with Type 2 Diabetes Mellitus Treated at Primary Health Clinics in Malaysia. Patient Preference and Adherence 2013 (7): 525-530
Wang X., Spandidos A., Wang H. and Seed B. 2012. Primerbank: A PCR Primer Database for Quantitative Gene Expression Analysis, 2012 update. Nucleic Acids Research 40 (D1): D1144-D1149
Zou Y., Jia X., Zhang A., Wang W.Z., Li S., Guo X., Kong Q.P., Zhang Q and Yao Y.G. 2010. The Mt-Nd1 and Mt-Nd5 Genes are Mutational Hotspots for Chinese Families with Clinical Features of LHON but Lacking the Three Primary Mutations. Biochemical and Biophysical Research Communications 399(2): 179-185
-- 0 --
