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Molecular Evolution of PPHLN1 Gene in Relation to HERV-M Element MOLECULAR BIOLOGY Jae-Won Huh1, Dae-Soo Kim2, Hong-Seok Ha1, Kung Ahn1, Yun-Ji Kim1, Ja-Rang Lee1, and Heui-Soo Kim1,2,* 1 Division of Biological Sciences, College of Natural Sciences, Pusan National University, Busan 609-735, Republic of Korea 2 PBBRC, Interdisciplinary Research Program of Bioinformatics, College of Natural Sciences, Pusan National University, Busan 609-735, Republic of Korea & PHYLOGENY LAB http://www.primate.or.kr Abstract Materials & Methods Results & Discussion 2 Novel Alternative Splicing of HERV-M related to PPHLN1 gene HERV-M (the human endogenous retrovirus M), related to the super family of HERV-K, has a Whole Genome sequence (Golden Path, hg16) methionine (M) tRNA primer-binding site and is located within the periphilin gene on human Reference Sequence (Feb. 27 2004) chromosome 12q12. HERV-M has been integrated into the periphilin gene as the truncated form, 5’LTR-gag-pol-3’LTR. PCR (polymerase chain reaction) and RT-PCR (reverse transcriptionHuman Tissues polymerase chain reaction) approaches were conducted to investigate its evolutionary origins. Interestingly, the insertion of various retroelements in a common ancestor genome may make dbEST clustering with template (Apr. 01 2004) Cancer cell lines different transcript variants in different species. In the case of the periphiline gene, human (10 variants) and mouse (2 variants) lineages show different transcript variants. Insertion of HERV-M Adult Stem cells RepeatMasker 3 The Structure of PPHLN1 Transcript variants (variant 1-3) could affect the protein coding region. Also Alusq/x (variant 4-9) and L1ME4a Monkey Tissue (mammalian-wide subfamilies of LINE-1) (variant10) in humans and SINE (short interspersed repetitive element) and RLTR15 (the mouse putative long terminal repeat) (variant 2) in mice could Total RNA isolation be driving forces in transcript diversification of the periphilin gene during mammalian evolution. Identification of retroelement related Gene The HERV-M derived transcripts (variant 1-3) were expressed in various human tissues, whereas it was not detected in crab-eating monkey and squirrel monkey tissues by RT-PCR amplification. mRNA subtraction Taken together, HERV-M seems to have been integrated into our common ancestor genome after the divergence of simians and prosimians, and then was actively expressed during hominoid Confirm with RT-PCR evolution...…………………………………………………………………………………… Introduction 1 Unidentified Gene New finding of Hybrid Genes with HERVs Results & Discussion 4 Evolution of HERV-M related alternative transcript 5 Expression profiling of HERV-M related alternative transcripts Insertion of Retroelements during the Evolution of PPHLN1 gene Genome Information Laboratory, Pusan National University, Republic of Korea