STUDI HOMOLOGI DAERAH TERMINAL-C HASIL TRANSLASI INSCRIPTO BEBERAPA GEN DNA POLIMERASE

• Main Author: MULYONO
• Format: Theses
• Language: Indonesia
• Subjects: Kimia
• Online Access: https://digilib.itb.ac.id/gdl/view/1663
• Institution: Institut Teknologi Bandung

DNA polymerase plays an important role in DNA replication process. <br /> Commonly the enzyme has three activities, i.e. 5\'--+3\' polymerase, <br /> 5\'-*3\' exonuclease and 3\'->5\' exonuclease which localized in three <br /> different domains. Orderly the polymerase activity lies on the Cterminal <br /> domain, followed by 3\'->5\' and 5\'-+3\' exonuclease activity <br /> domain on the N-terminal. Information concerning amino acid <br /> sequences on the C-terminal region is important for structure-function <br /> studies of the polymerase activity. Homology studies on DNA <br /> polymerases from different sources by observed on the sequence of the <br /> terminal amino acids have been conducted. C-terminal region of DNA <br /> polymerases (from GenBank i.e. NCBI) were compared which resulted <br /> in 2 conserved amino acid regions i.e. DPNLQNIP and QVHDELL. <br /> These regions which located on the C-terminal of DNA polymerase <br /> has one gap around 200 amino acids. One set of primer pairs were <br /> designed to amplify DNA fragment on the above region. Twelve out <br /> of fourteen of DNA samples, isolated from different organisms, were <br /> amplified by FP1 and PUR1 primers which resulted in 0.6 kb <br /> fragments. These fragments were cloned on pGEM-T vector using E. <br /> colt JM 109 as it\'s host. Restriction analysis on the recombinant <br /> plasmids suggested that the fragment were succesfully ligated. Further <br /> analysis based on nucleotide sequencing suggested that the fragments <br /> length were between 639 to 645 base pairs. All sequences were in <br /> frame with the published DNA polymerase. Inscripto translation and <br /> amino acids comparison among the fragments indicated that the <br /> sequences were relatively different. However, one out of three acidic <br /> amino acids that presumed has an important role on this region was <br /> very conserved as well. <br /> Comparison of the same region of DNA <br /> polymerases with the published data suggested that the organisms <br /> within a genus has specific pattern of amino acid sequences.