5) dn/ds ratio. / (dn/ds ratio) dn/ds ratio codon substitution model[3] 1) insulin GLP-1

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2 Gekko japonicus insulin proglucagon insulin GLP-1 GLP-2 mrna insulin proglucagon 1) (Gekko japonicus) -80 2) RNA cdna Total RNA ISOGEN (Nippon Gene) RACE cdna SMART RACE cdna Amplification Kit (BD Bioscience Clontech) 3 μg total RNA RT-PCR cdna 3μg total RNA M-MLV reverse transcriptase (Promega) 1/6-30 3) CLUSTAL X ve. 2.1 ORF default dn/ds ratio / (dn/ds ratio) dn/ds ratio codon substitution model[3] PAML4.7 CODEML Log likelihood Ratio Test default 4) CLUSTAL X (ver.2.1 ( org/clustal2/) ORF default 5)dN/dS ratio / (dn/ds ratio) dn/ds ratio codon substitution model[3] PAML4.7 ( gene.ucl.ac. uk/software/paml.html) CODEML Log likelihood Ratio Test default 5) mrna Oligo (dt) M-MLV Reverse Transcriptase 25ng/µL cdna PCR EF-1α Elongation Factor-1 alpha insulin proglucagon 1) insulin 1-1) proinsulin

3 Anguimorpha1 1 1 proinsulin cdna proinsulin 44-90% 45-90%(Anolis carolinensis) 49-95% insulin 60-92% % proinsulin 43% 50% 51%insulin 60% 58% 58% proinsulin 62% 62% 67%insulin 72% 74% 80% 10-30% 2 chain C-peptide insulin B-chain C 2 3 A-chain C Ser Lys 2 C Ile Glu Ser 3 (A7-B7 A20-B19 A6-A11) Cys PC 1-2) dn/ds ratio insulin / (dn/ds ratio) dn/ds ratio PAML4 7 CODEML 1 5% dn/ds ratio 1) 2) 3) 4) 5) 6) 6 4) 5) 6) 1 dn/ds ratio 1 PAML dn/ds ratio(=ω) insulin dn/ds ratio NCBI GenBank dn/ds ratio 2) proglucagon mrna 2-1) proglucagon mrna Class A/B mrna 2 cdna PCR PPG-A/-B Class A/B mrna ORF RT-PCR 3 Class A mrna PCR PPG-A (ORF 456 ) ORF (294 ) Class A mrna Class B mrna PPG-B (ORF 642 ) ORF (288 ) Class B mrna ORF (480 ) Class B mrna 2

2 proglucagon mrna DNA signal peptide 1 PPG-A 1 glucagon GLP-1 PPG-A 2 GLP-1 PPG-B 1 glucagon GLP-1 GLP-2 PPG-B 2 GLP-1 GLP-2 PPG-B 3 GLP-2 2-2) proglucagon RT-PCR proglucagon

PPG-A 1 (proglucagon A 1 PG-A 1 ) PPG-B 1 (PG-B 1 ) cdna PGDPs N/C PC2 PC1/3 exon 4 cdna cdna RT-PCR proglucagon exon 3 (PPG-A 2 /-B 2 /-B 3 ) Class A mrna PPG-A 2 PPG-A 1

4 2 proglucagon mrna DNA signal peptide 1 PPG-A 1 glucagon GLP-1 PPG-A 2 GLP-1 PPG-B 1 glucagon GLP-1 GLP-2 PPG-B 2 GLP-1 GLP-2 PPG-B 3 GLP-2 2-2) proglucagon RT-PCR proglucagon Class A mrna PPG-A 1 /-A 2 PPG-A1 PPG-A 2 Class B mrna PPG-B 1 /-B 3 PPG-B 3 PPG-B 3 PPG-B 2 /-B 3 PPG-B 3 PPG-B 3 2-3) proglucagon PPG-A/-B PPG 1 10 proglucagon mrna PPG-A 1 (proglucagon A 1 PG-A 1 ) PPG-B 1 (PG-B 1 ) cdna PGDPs N/C PC2 PC1/3 exon 4 cdna cdna RT-PCR proglucagon exon 3 (PPG-A 2 /-B 2 /-B 3 ) Class A mrna PPG-A 2 PPG-A 1 Unidentata ( ) 5 PPG-A 1 /-A 2 PPG-A 1 PPG-A 2 PPG-A 1 Class B mrna PPG-B 2 PPG-B 1 Unidentata 5 PG-B 1 /-B 2 PPG-B 1 PPG-B 2 PPG-B 3 3) insulin GLP-1 GLP-2 mrna insulin adiponectin

5 3 ( ) Mawaribuchi S, Musashijima M, Wada M, Izutsu Y, Kurakata E, Park MK, Takamatsu N, and Ito M. (2017) Molecular Evolution of Two Distinct dmrt1 Promoters for Germ and Somatic Cells in Vertebrate Gonads. Molecular biology and evolution 34(3): DOI: /molbev/msw273 Yamagishi G, Yoshida A, Kobayashi A, and Park MK (2016) Molecular characterization of insulin from squamate reptiles reveals sequence diversity and possible adaptive evolution. Gen. Comp. Endocrinol. 225(): DOI: /j.ygcen Otake S. and Park MK (2016) Expressional changes of AMH signaling system in the quail testis induced by photoperiod. Reproduction 152(5): DOI: /REP Yamagishi G, and Park MK (2016) Lipid accumulation and adipose tissue-like gene expression in the liver of the Japanese gecko (Gekko japonicus). The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society of Japan, Nov , Okinawa Convention Center (Okinawa, Ginowan), Abstract p404. Kaiya H, Park MK, Kanagawa K, and Miyazato M (2016) Identification of ghrelin and its receptor in Japanese gecko, Gekko japonicus. The joint meeting of the 22nd International Congress of Zoology and the 87th Meeting of the Zoological Society of Japan, Nov , Okinawa Convention Center (Okinawa, Ginowan), Abstract p p p proglucagon mrna p insulin p71 Oatake S, and Park MK (2015) Expression changes of genes related to the AMH signaling systems during the progression of spermatogenesis in the Japanese quail JMS p51 Kurakata E, and Park MK (2015) Characterization of preproglucagon mrna variants encoding only glucagon in Gekkota JMS p63 Yamagishi G, and Park MK 2015 Molecular evolution and tissue distribution of glucose-6-phosphatase catalytic subunit (G6PC) in reptiles JMS p40 Kurakata E, Suzuki Y, Yoshida A, Kobayashi A, and Park MK (2014) Molecular characterizations and expression analysis of preproglucagon mrna variants specific to reptiles. In Abstract of JSCE 2014 and ISAREN 2014, pp 97. (Session 5: Metabolism, The 8th International Symposium on Amphibian and Reptilian Endocrinology and Neurobiology (ISAREN 2014), November 7-9, 2014, National Institute for Basic Biology (Aichi, Okazaki). Yamagishi G, Yoshida A, Kobayashi A, and Park MK (2014) Molecular characterization of highly divergent insulins from squamates. In Abstract of JSCE 2014 and ISAREN 2014, pp98. (Session 5: Metabolism, The 8th International Symposium on Amphibian and Reptilian Endocrinology and Neurobiology (ISAREN 2014), November 7-9, 2014, National Institute for Basic Biology (Aichi, Okazaki). Yamagishi G, and Park MK (2014)

6 Molecular characterization of squamate insulins and its receptors, and their evolutionary implication. In Abstract of JSCE 2014 and ISAREN 2014", pp155. (Joint Meeting of the 39nd Annual Meeting of the Japanese Society of Comparative Endocrinology (JSCE) and the 8th International Symposium on Amphibian and Reptilian Endocrinology and Neurobiology (ISAREN 2014), Nov. 7-9, 2014, National Institute for Basic Biology (Aichi, Okazaki). Otake S, and Park MK (2014) Expression analysis of TGF-β superfamily members in the adult testis of the Japanese quail, In Abstract of JSCE 2014 and ISAREN 2014, pp151. (Joint Meeting of the 39nd Annual Meeting of the Japanese Society of Comparative Endocrinology (JSCE) and the 8th International Symposium on Amphibian and Reptilian Endocrinology and Neurobiology (ISAREN), Nov. 7-9, 2014, National Institute for Basic Biology (Aichi, Okazaki). [ ] 6 (2016) (pp84-100) Park MK (2016) Glucagon family, In Handbook of Hormones: Comparative Tsutsui, Elsevier, 646 (pp , +5 Park MK (2016) Gastric Inhibitory Peptide, In Handbook of Hormones: Comparative Tsutsui, Elsevier, 646 (pp , +7 Park MK (2016) Glucagon, In Handbook of Hormones: Comparative Endocrinology for Basic and Clinical Research, Ed by Y. Takei, H. Ando and K. Tsutsui, Elsevier, 646 (pp , +6 Park MK (2016) Glucagon-like peptide-1, In Handbook of Hormones: Comparative Tsutsui, Elsevier, 646 (pp , +7 Park MK (2016) Glucagon-like peptide-2, In Handbook of Hormones: Comparative Tsutsui, Elsevier, 646 (pp , +7 (1) PARK, MIN KYUN (2) (3)

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