NUCLEOTIDE MUTATIONS ANALYSIS OF MITOCHONDRIAL GENOME OF TWO INDONESIAN HUMAN INDIVIDUALS AND SEVERAL INDIVIDUALS FROM GENBANK

• Main Author: NGILI (NIM : 30507007); Ketua tim promotor : Achmad Saifuddin Noer,Ph.D; Anggota tim pembi, YOHANIS
• Format: Dissertations
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/17645
• Institution: Institut Teknologi Bandung

Research related to the analysis of DNA and its application in various fields of science and technology is the leading branch of science because it helps answer the basic human problems of the present and future. Hipervariabel nature of <br /> <br /> <br /> mtDNA has been associated with the comparison of nucleotide sequences and mtDNA mutations among individuals, ethnic and age. However, hypervariable mutation occurred in mtG compared with rCRS (revised-Cambridge Reference Sequence) and hypervariable in various tissues of different origins such as layers entoderm, mesoderm, and ectoderm in certain individuals has not been reported previously. Also, it will compare nucleotide mutations in coding regions of genes, <br /> <br /> <br /> and seen new mutations or mutations that have not been reported in MITOMAP. <br /> <br /> <br /> In this dissertation nucleotide sequence of the mitochondrial DNA from two human individuals Indonesia, which has not been reported previously, have successfully sequenced the nucleotide sequence obtained through protocol <br /> <br /> <br /> purification of DNA from tissues and repli-G. Designing primers for amplification performed 10 fragments (fragments of A, B, C, D, E, F, G, H, I, and M) with respect to aspects of efficiency. Mutation analysis is done using a variety of data processing programs such as DNASTAR: Seqman, MegAlign, Editseq; H-Man; Excel Spreadsheet Program v.3 and v.4. The results of mutation analysis showed a <br /> <br /> <br /> point mutation in almost all the fragments with different proportions. Nucleotide mutation in a single individual among tissue showed a similar pattern in both the D-loop region (control region) and in the coding region of the gene. This suggests that the mutation occurred in one individual is to be monomorphic. <br /> <br /> <br /> Variant mutations that occur in the human mitochondrial genome Individual A compared with rCRS as follows. Two variants of the 12S rRNA, one variant of the 16S rRNA, one variant in the region ND1, two variants in the ND2 region, one variant of the local interregion (int), three variants of the CO1 region, one variant in the CO2 area, five variants in the ATP6 region, one variant of CO3 region, two <br /> <br /> <br /> variants in the ND3 region, two variants in the region of ND4, three variants in the ND5 region, two variants on CYB area, and five variants of the D-loop region. <br /> <br /> <br /> For individual B, mutation variants include one variants on the 12S rRNA region, one variant of the 16S rRNA region, one variant of the ND1 area, one variant in the ND2 region, one variant in the int region, one variant in the of CO2 region, one variant in the ATP6 region, one variant in CO3, one variant in the region of ND3, two variants in the ND4 region, two variants of the ND5 region, three variants of the CYB area, and six variants of the D-loop region. <br /> <br /> <br /> In addition, the results of this study also obtained nine mutation variants that have not been reported in MITOMAP from the Indonesian population sequences. Varian mutation that has not been reported is T650c, A3873g, C5213g,A6863g, A7894g, A8652g, A8653g, G10373c, and C15211t. Most types of mutations that occur in the mitochondrial genome is transition substitution mutations. Variant mutations show differences against rCRS is on areas such as the D-loop, rRNA, ND2, CO1, ATP6, ND3, ND4, ND5, CYB. <br /> <br /> <br /> From the results of this research, and the development of new lines of inquiry that can be generated from this study is to further study the isolation and amplification of whole genome amplification methods can be developed so that the mitochondrial genomes are cheaper and more efficient. Other research that can be developed is a study to determine new mutations (which have not been reported in <br /> <br /> <br /> MITOMAP), especially those related to disclosures variant mutation in the coding region of the gene and the expression of biochemical and clinical manifestations <br /> <br /> <br /> that occur in mitochondrial disease. By creating a database of mutations associated with a particular disease allows more quickly determine the status of a person's illness. In addition, the approach taken in this study can be used as a <br /> <br /> <br /> reference in the study of other mutations associated with mitochondrial disease is suspected, especially for Indonesian people of various ethnic group.