Real-time RT-PCR

1. [Figure 3. Expression analysis of alternative transcripts in various human tissues including different brain tissues.] 8% Human Real-time RT-PCR Gorilla Orangutan LTR33 Gibbon Old World monkeys New World monkeys Prosimians Million years 70 60 50 40 30 20 10 Dual coding capacity of MOBP gene derived by the insertion of retroviral LTR33 element Yu-Na Noh1, Jae-Won Huh1, Dae-Soo Kim2, Kung Ahn1, and Heui-Soo Kim1,2 1.Division of Biological Sciences, College of Natural Sciences, Pusan National University 2.PBBRC, Interdisciplinary Research Program of Bioinformatics, College of Natural Sciences, Pusan National University RESULTS & DISCUSSION ABSTRACT We investigated the association of LTR element in transcript variant of MOBP gene. LTR33 belonging to the ERV1 family contributes to the change of coding region of MOBP gene by te production of splicing variant. Real-time RT-PCR analysis using different human tissues shows the dominant expression patterns in AD brain, adult whole brain, spinal cord, and testis tissues. Interestingly, comparative analysis of expression pattern using corresponding tissues between human and rhesus monkey shows the different expression patterns. To investigate the evolutionary conservation of LTR33 element of the MOBP gene, various primate DNA samples were used for the PCR amplification and sequencing. Their splicing donor site (GT) are completely conserved and the number of tandem repeated element which could be translated are increased during the evolution of hominoid primates. Taken together, integration event of LTR33 element could affect the coding capacity and expression in different tissues by changing the coding sequences during the primate evolution. [Figure 1. Four different alternative transcripts of MOBP gene] INTRODUCTION [Figure 2. Multiple alignment of MOBP gene.] [Structure of Retrovirus] [Retrovirus life cycle] [HERV Structure] [Genome component parts] *HERV: Human Endogenous Retrovirus [Figure 4.Comparative analysis of MOBP gene transcripts between Human and Rhesus Monkey] MERERIALS & METHODS PCR Primate DNA MOBP Gene MOBP_LTR33/GAPDH [Figure 5. Real-time RT-PCR analysis of MOBP gene transcript] Gene Cloning Bio Informatics Amplified Plasmid DNA REFERENCES [Figure 6.Integration lineage of LTR33 element during the Primate evolution.] 1.Yoichi Yamamoto et al. 1994,Cloning and Expression of Myelin-associated Oligodendrocytic Basic Protein, The Journal of Biological Chemistry,Vol.269:31725-31730 2. HOLZ A. and SCHWAB M. E.,1997,Developmental Expression of the myelin gene MOBP in the rat nervous system, Journal of Neurocytology,26:467-477 www.primate.or.kr