Molecular detection of SVA-derived transcripts

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Molecular detection of SVA-derived transcripts in human MRGPRX3 gene Yu-Na Noh1, Jae-Won Huh2, Dae-Soo Kim3, Hong-Seok Ha1, Kung Ahn1,Hwan-Gue Cho4 and Heui-Soo Kim1 1. Division of Biological Science, College of Natural Sciences, Pusan National University, Busan 609-735 2. National Primate Research Center (NPRC), KRIBB, Cheongwon 363-883 3. Korea Bioinformation Center, KRIBB, Daejeon 305-806 4. Dept. of Computer Sciences, Pusan National University, Busan 609-735, Republic of Korea Abstract MRGPRX3 [MAS-related GPR (G protein-coupled receptor) member X3] gene located on chromosome 11p15.1 region, and MRGPRX3 gene were as known as sensory neuron specific receptors that detect painful stimuli and associated with modulation of nociception. MRGPRX3 gene has alternative transcripts which are SVA-derived transcripts and original transcripts. Using experimental approaches and computational screening, we identified three SVA-derived and two original transcripts. Intriguingly, three SVA-derived transcripts are expressed specifically in testis tissues. However, original transcript is corresponding to transcribe ubiquitously in several tissues. Integration lineage of SVA element was analyzed by PCR with various primate DNA samples. The results indicated that SVA element in MRGPPRX3 gene has been integrated into our common ancestor genome after the divergence of human and chimpanzee lineage. Taken together, integration event of SVA element in MRGPRX3 gene during the primate evolution seems to contribute the diversity of human transcriptome by providing testis specific promoter and following alternative transcripts. Introduction Results and Discussion [ Figure 1. Alternative transcripts of MRGPRX3 gene ] [Structure of Retrovirus] [Cycle of Retrovirus] [Figure 2. Integration lineage of SVA element during the Primate evolution.] [Structure of SVA ( SINE (short interspersed element), VNTR(variable number of tandem repeat) and Alu)] [Figure 3. RT-PCR analysis of MRGPRX3 gene transcript] [Amplification dynamics of the SVA family of retroposons in primates. Putative evolutionary history of the SVA lineage in primate genomes.] [Figure 4. Sequence alignment of MRGPRX3 gene transcript] Materials and Methods [Figure 5. Alternative transcripts of MRGPRX3 gene by SVA] PCR Primate DNA MRGPRX3 Gene References Hui Wang et al., 2005, SVA Elements: A Hominid-specific Retroposon Family, Journal of Molecular Biology, Vol.354:994–1007 Satoshi Makino et al., 2007, Reduced Neuron-Specific Expression of the TAF1 Gene Is Associated with X-Linked Dystonia-Parkinsonism, The American Journal of Human Genetics, Vol.80, 393-406 Ryan E. Mills et al., 2007, Which transposable elements are active in the human genome?, TRENDS in Genetics, Vol.23, 183-191 Gene Cloning Bio Informatics Amplified Plasmid DNA www.primate.or.kr