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1 26 MHC I
2
3 major histocompatibility complex; MHC MHC MHC T (Klein, 1986) MHC (Kelley, 2005) (Zinkernagel, 1979) MHC MHC (Fernando, 2008) MHC MHC-B (Kaufman, 1999; Hosomichi, 2006; Chaves, 2009; Ye, 2012; Wang, 2012; Eimes, 2013) 3 MHC (Ekblom, 2011; Moon, 2011; Taniguchi, 2014) MHC MHC MHC 1
4 (Sommer, 2005) (Ezaki, 2013) (Frankham, 2002) (Reed, 2001) MHC MHC MHC MHC IUCN IA (IUCN, 2014) ,000 2,
5 ( DNA 26 D 9 (Yamamoto, 2000) (16 4 ) (80 33 ) (8 3 ) (, 2012) MHC MHC 3
6 MHC MHC MHC II COL11A2-MHC IIA/IIB-BRD2 6 9 ( 辻, 2015) MHC I MHC I 4
7 PCR 1 A11B F/R A11B BRD2 F/R BRD2 A11B A11B 3 BRD2 ( 辻, 2015) TAP1 exon7 F/R TAP1 7 PCR ( 辻, 2015) PCR Taniguchi (2014) PCR KOD FX Neo Buffer (TOYOBO ) 7.5 dntp ( 2.5mM) 3 ( 50 M) 0.75 KOD FX Neo (TOYOBO ) 0.3 DDW DNA % 30 2 Q17G A11B F/R A11B BRD2 F/R Q13C TAP1 exon7 F/R K8A K6J Q13C 5
8 6 1.PCR
9 2. 7
10 5 3 K6J 3 T69H V66E Z34D MHC I F/R N49F MHC I F/R Z34D F/R DNA DNA Q17G BamHI K8A Q13C EcoRI XhoI EcoRI pbluescriptii Competent Quick DH5 TOYOBO DNA Mini PlusTM Plasmid DNA Extraction System VIOGENE ( ) BLAST ( GENETYX version 11 (Software Development) PCR 4 (K6J, T69H, V66E, N49F) 5 (K6J F/R, V66E S/A, T69H F/R, V66E F/R, MHC I F/R) PCR PCR PCR % 30 8
11 MHC MHC 8 MHC I BRD2 (2015) Moon (2011) Kaufman (1999) Chaves (2009) Ye (2012) Wang (2012) Eimes (2013) Shiina (2004) MHC I 8 DNA ( 1) II DNA 100kb Illumina HiSeq 2500 Velvet de novo BLAST MHC 9
12 1. MHC BRD2 DMA DMB1 DMB2 MHC I(UAA1) TAP1 TAP2 MHC I(UAA2) EcoRI B E X BamHI EcoRI XhoI DNA A11B* 辻, (2015) 10
13 MHC I MHC I MHC ( 2)BRD2 Q17G TAP1 Q13C Q17G BRD2 DMA DMB2 Q13C DMB2 MHC I(UAA1) TAP1 Q13C K8A K6J K8A Q17G Q13C K6J TAP2 K6J T69H V66E Z34D MHC I(UAA2) PCR V66E Z34D N49F 90kb BRD2 DMA DMB1 DMB2 UAA1 TAP1 TAP2 UAA2 T69H EcoRI N49F EcoRI ( 1, ) TAP2 UAA2 11
14 UAA2 4 5 PCR ( 2) K6J F/R V66E S/A PCR N49F T69H F/R PCR N49F V66E F/R PCR N49F V66E F/R V66E T69H MHC I F/R PCR K6J N49F N49F T69H V66E UAA2 K6J F/R T69H F/R ( 1, ) MHC I MHC I ( 3) I BRD2 - DMA - DMB1 - DMB2 - UAA1- TAP1 - TAP2 - UAA2 DMB2 TAP1 MHC I ( ) BRD2 TAP2 TAP2 UAA2 TAP2 12
15 2. PCR K6J T69H V66E N49F K6J F/R V66E S/A T69H F/R V66E F/R MHC-I F/R PCR 100bp (GeneDireX ) 13
16 3. MHC TAP1 TAP2 MHC I 14
17 MHC I MHC I 1 MHC I MHC I MHC I II Illumina HiSeq ,944, Gb ( II 辻 (2015) ) I 15 ( 1) I 80kb 57.5kb 11 ( S1) 8 DMA DMB1 TAP1 TAP2 UAA1 UAA2 MHC I 15
18 MHC I 90kb 8 MHC I ( 1) BRD2 - DMA - DMB1 - DMB2 - UAA1 - TAP1 - TAP2 - UAA2 ( 3) MHC I TAP1 TAP2 TAP1-TAP2 ( 3) TAP2 MHC I TAP2 MHC I 1 MHC I MHC IIB (Shiina, 2004) UAA2 MHC I UAA2 ( 1) TAP2 MHC I TAP C4 MHC I 1 ( 3) 16
19 MHC I TAP MHC I (UAA2-5) (, 2015) UAA3 12kb UAA4 TAP MHC I MHC I 2 MHC I (UAA1 UAA2) Velvet MHC I MHC I 90kb II 40kb ( 辻, 2015) MHC MHC MHC MHC I 17
20 MHC I (Aldridge, 2006; Garrigan, 2001) 6 II 9 ( 辻, 2015) I 18
21 (Ciconia boyciana) MHC MHC MHC MHC MHC MHC II 6 MHC I PCR 8 90kb MHC I BLAST BRD2 - DMA - DMB1 - DMB2 - MHC I(UAA1) - TAP1 - TAP2 - MHC I(UAA2) PCR UAA2 10kb MHC BRD2 19
22 TAP2 TAP2 UAA2 MHC TAP2 80kb 57.5kb 11 MHC 20
23 21
24 Aldridge, B. M., L. Bowen, B. R. Smith, G. A. Antonelis, F. Gulland, J. L. Stott Paucity of class I MHC gene heterogeneity between individuals in the endangered Hawaiian monk seal population. Immunogenetics, 58, , 2006 Eimes, J. A., K. M. Reed, K. M. Mendoza, J. L. Bollmer, L. A. Whittingham, Z. M. Bateson, P. O. Dunn Greater prairie chickens have a compact MHC-B with a single class IA locus. Immunogenetics, 65, , 2013 Ekblom, R., J. Stapley, A. D. Ball, T. Birkhead, T. J. Slate Genetic mapping of the major histocompatibility complex in the zebra finch (Taeniopygia guttata). Immunogenetics, 63, , 2011 Ezaki, Y., Y. Ohsako, S. Yamagishi Re-introduction of the oriental white stork for coexistence with humans in Japan. Global Re-introduction Perspectives, 85-89, 2013 Fernando, M. M. A., C. R. Stevens, E. C. Walsh, P. L. D. Jager, P. Goyette, R. M. Plenge, T. J. Vyse, J. D. Rioux Defining the Role of the MHC in Autoimmunity: A Review and Pooled Analysis. PLoS Genetics, 25, ,
25 Frankham R., J. D. Ballou, D. A. Briscoe Introduction to conservation genetics. Cambridge University Press, 2002 Garrigan, D., P. W. Hedrick Class I MHC polymorphism and evolution in endangered California Chinook and other Pacific salmon. Immunogenetics, 53, , 2001 IUCN The IUCN Red List of Threatened Species. Version Kaufman, J., S. Milne, T. W. Gobel, B. A. Walker, J. P. Jacob, C. Auffray, R. Zoorob, S. Beck The chicken B locus is a minimal essential major histocompatibility complex. Nature, 401, , 1999 Kelley, J., L. Walter, J. Trowsdale Comarative genomics of the major histocomatibility complexes. Immunogenetics, 56, , 2005 Klein, J. Antigen-Major Histocompatibility Complex-T cell receptors: Inquiries into the immunological menage a trois. Immunologic Research, 5, ,
26 MHC., 1-70, 2015 Moon, D. A., S. M. Veniamin, J. A. Parks-Dely, K. E. Magor The MHC of the duck (Anas platyhynchos) contains five differentially expressed class I genes. The Journal of Immunology, 175, , 2005,,., 2, 57-62, 2012 Reed, D. H., R. Frankham How closely correlated are molecular and quantitative measures of genetic variation? A meta-analysis. Evolution, 55, , 2001 Shiina, T., S. Shimizu, K. Hosomichi, S. Kohara, S. Watanabe, K. Hanzawa, S. Beck, J. K. Kulski, H. Inoko Comparative genomics analysis of two avian (quail and chicken) MHC regions. The Journal of Immunology, 172, , 2004 Sommer, S. The importance of immune gene variability (MHC) in evolutionary ecology and conservation. Frontiers in Zoology, 20, 2-16,
27 Taniguchi Y., K. Matsumoto, H. Matsuda, T. Yamada, T. Sugiyama, K. Homma, Y. Kaneko, S. Yamagishi, H. Iwaisaki Structure and polymorphism of the major histocompatibility complex class II region in the Japanese Crested Ibis, Nipponia nippon. PLoS One, 9, e108506, 2014 辻 MHC II., 1-70, 2015 Yamamoto, Y., K Murata, H. Matsuda, T. Hosoda, K. Tamura, J. Furuyama Determination of the complete nucleotide sequence and haplotypes in the D-loop region of the mitochondrial genome in the oriental white stork, Ciconia boyciana. Genes and Genetic Systems, 75, 25-32, 2000 Ye, Q., K. He, S. Wu, Q. Wan Isolation of a 97-kb minimal essential MHC B locus from a new reverse-4d BAC library of the golden pheasant. PLoS One, 7, e32154, 2012 Wang, B., R. Ekblom, T. M. Strand, S. Portela-Bens, J. Hoglund Sequencing of the core MHC region of black grouse (Tetrao tetrix) and comparative genomics of the galliform MHC. BMC Genomics, 25
28 Zinkernagel, R. M., P. C. Doherty MHC-restricted cytotoxic T cells: studies on the biological role of polymorphic major transplantation antigens determining T-cell restriction-specificity, function, and responsiveness. Advance in Immunology, 27, ,
Niigata University Ⅰ. 研究の背景日本の特別天然記念物であり 希少動物であるトキ コウノトリの保全が国家的プロジェクトとして進められています このような個体数の極めて少ない希少動物集団の保全においては 集団内の遺伝的多様性を維持することが重要な課題となっています 主要組織適合遺伝
Niigata University 平成 29 年 4 月 3 日 希少動物であるトキ コウノトリの保全および鳥類の進化を探る上での重要な発見 ーニワトリ ( キジ目 ) の MHC 領域のゲノム構造は鳥類の典型ではないー 希少動物であるトキ コウノトリの保全においては 遺伝的多様性の把握と維持が重要な課 題となっています 本学研究推進機構朱鷺 自然再生学研究センターの祝前博明特任教授 本学農学部の山田宜永教授らの研究グループは
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