Prediksi DNA Primer gen PGC-1α cecak (Hemidactylus platyurus) dengan metoda phylogenetic, multiple alignment, dan qPCR

Abstract

The role of genes in tissue regeneration process of house gecko (Hemidactylus platyurus) tail is very important to study. However, the PGC-1α gene that plays a role in mitochondrial biogenesis in the tissue regeneration process of house gecko tail has not been studied before. The phylogenetic method can be used to track the determinant species closest to kinship. Multiple alignment method is software that can align the conserved gene sequences from several species that have sequence identities highly. The phylogenetic analysis is used to track the determinant species closest to kinship. Basic Local Alignment Software Tools (BLAST) is used to analyze the identity of gene sequences from several species. Alignment the conserved sequences of gene was using to multiple alignment methods in MEGA7 software. Primary DNA predicted of PGC-1α gene was made using Primer3 software that based on gene sequences conserved from MEGA7 multiple alignment process. The specification of gene expression was tested by Real Time PCR. The results of phylogenetic analysis the species closest to H. platyurus are Gecko japonicus. BLAST results and multiple alignments obtained by sequences as a basis for the design of primary DNA. The primary design of the DNA predicted PGC-1α H.platyurus gene was successfully amplified with Real Time PCR and produced one peak, which indicates the specific gene expression. Predicted primary DNA PGC-1α gene of Hemidactylus platyurus that has not been previously studied, can be designed using phylogenetic to determinant species closest to kinship and multiple alignment methods.