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1 37 5 Vol. 37, No ACTA HYDROBIOLOGICA SINICA Sep., doi: 0.754/ GS FLX ESTs 曾聪, 2 高泽霞罗伟刘肖莲王卫民 (.,,, ; 2., ) CHARACTERISTICS OF MICROSATELLITES IN BLUNT SNOUT BREAM (MEGALOBRAMA AMBLYCEPHALA) EST SEQUENCES USING 454 FLX ZENG Cong, GAO Ze-Xia, 2, LUO Wei, LIU Xiao-Lian and WANG Wei-Min (. Key Lab. of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Key Lab. of Freshwater Animal Breeding, Ministry of Agriculture, College of Fishery, Huazhong Agricultural University, Wuhan , China; 2. Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan , China) : ; EST-SSR; Key words: Megalobrama amblycephala; EST-SSR; Characteristics 中图分类号 : Q 文献标识码 : A 文章编号 : (203) (Simple Sequence Repeat, SSR), [], SSR,, (Expressed Sequence Tag, EST) (EST-SSR),,, EST, EST Sanger,, PCR,,, (Megalobrama amblycephala Yih), (Cypriniformes), (Cyprinidae), (Megalobrama),,, 955,,, 20 60,,,, 985,,,, 999 [2], NCBI ( nlm.nih.gov) 60, EST-SSR 收稿日期 : ; 修订日期 : 基金项目 : (CARS-46-05); (320988) ( 20PY PY066); 作者简介 : (987 ),, ; ; congzeng@live.cn 通信作者 :,, ; wangwm@mail.hzau.edu.cn

of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, Key Lab.

2 5 : 454 GS FLX ESTs GS FLX, ESTs SSRs, EST-SSRs, EST-SSRs. 2009,, 2,,, 80RNA 454 [3].2 EST [3] EST, (Ctenopharyngodon idellus) (Cyprinus carpio) (Carassius auratus) NCBI EST.3 SSR, Msatcommander ( unigene SSR, SSR SSR 2, 2 SSR 50 bp,, EST SSR, 50 bp 2 2. EST-SSRs 454 GS FLX Titanium, EST, 4 bp, unigene 00477, contig singlet, unigene 47.7 Mb GenBank dbest, 20 Sanger EST bp , EST 3.90%.24% 0.33%, 97% SSR,, SSR, EST 3.93 kb SSR,, EST-SSR, ( ) 2.2 EST-SSR EST-SSR,,,,, 24 bp, 80% EST-SSR,, 2 bp ; 36 bp, 95% EST-SSR,, 24 bp,, 36 bp (A)n;, CG/CG 24 bp, AC/GT AG/CT AT/TA, AC ;, AAG, ACG/CGT, CCG/CGG 24 bp, ACT, 36 bp;, AGAT/ATCT, CCGG/GGCC ; 3046 EST-SSRs, EST, 2 322, ( ),,, EST, W(Shapiro-Wilk) D(Kolmogorov-Smirnov) ( ) 0.05 (P<0.0), EST-SSR Fig. Length of Repeat motif in EST-SSR of M. amblycephala

EST-SSRs 454 GS FLX Titanium, 409706 EST, 4 bp, unigene 00477, 26802 contig 73675 singlet, unigene 47.7 Mb GenBank dbest, 20 Sanger EST 2 24 36 bp 3046 6 30, EST 3.90%.24% 0.33%, 97% SSR,, SSR, EST 3.

3 984 37

4 5 : 454 GS FLX ESTs 985 EST EST-SSR 2 32 bp, 2 bp, 6007, ; 5 bp, 343, 322 bp, bp SSR SSR 84.64%, bp.93%, 30 bp 3.43% 2, EST-SSR 3, 3, 2 23, 23, EST-SSR, 9405, 73.24%, 2 6, ; 3322, 25.87%, ; 4,, 0.89% (65.39%), CG/CG ( 0.55%);, ATC/GAT, SSR 22.27%;, AAAT/ ATTT ; AAAAC/ AAAAAC/GTTTTT AATGAG/ CTCATT,, EST-SSR,,, 3 EST SSR EST 3.89%, NCBI EST (C. idellus) (C. carpio) (C. 2.3 EST-SSR NCBI (C. idellus) (C. auratus) SSR, EST SSR, (Haliotis discus carpio) (C. auratus) EST ESTs hannai) 6.25% [4] (Penaeus monodon) 5247, 5455, 35098, 3.7% [5] 鲀 (Fugu rubripes).5% [6] 594 EST 83 SSR, (Ictalurus punctatus).2% [7], (Miichthys miiuy) 8.29% [8] (Paralichthys olivaceus) 33.24%; 750, 29.78%; 3453, 7.95% [9] (Pseudosciaena crocea) 6.75% [0] 33.48% 2 (Apostichopus japonicus) 6.48% [] (Pagrosomus major) EST-SSR EST-SSR, EST-SSR, , 4.00% [2] (Argopectens irradias) 3.9% [3] (Crassostrea gigas) 3.63% [4] (Penaeus chine) 2.2% [5] (Mizuhopecten yessoensis).78% [6] (Chlamys farreri).6% [7] EST-SSR, SSR [8], EST-SSR, A/T (95%); SSR,,, CA/GT, [9] EST-SSR, A/T G/C (>95%), poly(a/t) Alu LINE- ; ( 2) 2 EST-SSR Fig. 2 Distribution of EST-SSR repeats in M. amblycephala,, AC/GT,,

idellus) (C. auratus) SSR, EST SSR, (Haliotis discus carpio) (C. auratus) EST ESTs hannai) 6.25% [4] (Penaeus monodon) 5247, 5455, 35098, 3.7% [5] 鲀 (Fugu rubripes).

5 表 2 草鱼鲤鲫和团头鲂的不同重复基元的 EST-SSR 比较 Tab. 2 Compare of different repeats in EST-SSR among Ctenopharyngodon idellus, Cyprinus carpio, Carassius auratus and M. amblycephala C. idellus C. carpio C. auratus M. amblycephala of predomination of predomination of predomination of predomination Mononucleotide Dinucleotide Trinucleotide Tetranucleotide Pentanucleotide Hexanucleotide 59.76% 2.83% 2.73% 2.3%.94% 0.42% A/T AC/TG ATC/GAT AAAT/ATTT AAAAC/GTTTT ACCTCC/GGAGGT 98.73% 6.5% 27.22% 38.7% 27.03% 2.50% 4.04% 25.39% 8.36%.84% 2.0%.27% A/T AC/GT AAT/ATT AAAT/ATTT AAAAT/ATTTT ACACAG/CTGTGT 96.8% 49.23% 8.57% 35.06% 22.92% 27.59% 38.4% 24.58% 7.30% 5.0% 3.26%.45% A/T AC/GT AAT/ATT AAAT/TTTA AAAAT/TTTTA ACACAG/CTGTGT 89.76% 57.47% 23.42% 29.99% 29.5% 8.08% 2.2% 27.00% 23.95% 2.36% 4.80%.67% A/T AC/GT ATC/GAT AAAT/ATTT AAAAC/GTTTT AAAAAC /GTTTTT AATGAG/CTCATT 97.6% 65.39% 22.27% 25.52% 27.60% 3.02% [] ;,,,,,, [8,9], CCG/CGG, Gabor, et al. CCG/CGG [20],, CCG/CGG,, ;,, 76,,,, EST-SSR, 6, SSR,, SSR [2] [22], ( ) EST-SSR, 30 bp,,,,, [23] Temnykh, et al. [24], SSR 20 bp,,, 2 20 bp, 2 bp,, AC, SSR, 39 [25 27], bp (69.23%), AC/TG 2 (53.85%),

amblycephala of predomination of predomination of predomination of predomination Mononucleotide Dinucleotide Trinucleotide Tetranucleotide Pentanucleotide Hexanucleotide 59.76% 2.83% 2.73% 2.3%.94% 0.

6 5 : 454 GS FLX ESTs 987 EST-SSR SSR, EST-SSR,, EST-SSR [28], EST-SSR,, [29 30], EST-SSR : [] Powell W, Machray G C, Provan J. Polymorphism revealed by simple sequence repeats [J]. Trends in Plant Science, 996, (7): [2] Li S F. Introduction of the fish good breed Pu-jiang No. blunt snout bream [J]. China Fisheries, 200, : 52 [.., 200, : 52] [3] Gao Z, Luo W, Liu H, et al. Transcriptome analysis and SSR/SNP markers information of the blunt snout bream (Megalobrama amblycephala) [J]. PLoS One, 202, 8(2): e42637 [4] Yuan Z Y, Zhang L L, Zhang G F. Bioinformatic date mining of EST- tandem repeats of the Haliotis discus hannai [J]. Marine Science Bulletin, 200, 29(): [,,. (Haliotis discus hannai)est., 200, 29(): 59 64] [5] Maneeruttanarungroj C, Pongsomboon S, Wuthisuthimebavee S, et al. Development of polymorphic expressed sequence tag derived microsatellites for the extension of the genetic linkage map of the black tiger shrimp (Penaeus monodon) [J]. Animal Genetics, 2006, 37(4): [6] Edwards Y J, Elgar G, Clark M S, et al. The identification and characterization of microsatellites in the compact genome of the Japanese puffer fish, Fugu rubripes: Perspectives in functional and comparative genomic analyses [J]. Journal of Molecular Biology, 998, 278(4): [7] Serapion J, Kucuktas H, Feng J N, et al. Bioinformatic mining of type microsatellites from expressed sequence tags of channel catfish (Ictalurus punctatus) [J]. Marine Technology, 2004, 6(4): [8] Sun D Q, Sun Y N, Wang R X, et al. The characteristic analysis of microsatellites from ests in miichthys miiuy [J]. Acta Hydrobiologica Sinica, 20, 35(5): [,,,. EST., 20, 35(5): ] [9] Chen S B, Gong L, Liu H J. Analysis of SSR information in EST resources of Japanese flounder (Paralichthys olivaceus) [J]. Journal of Northeast Agricultural University, 200, 4(0): [,,. EST SSR., 200, 4(0): 82 86] [0] Ye H, Wang X Q, Gao T X, et al. EST-derived microsatellites in Pseudosciaena crocea and their applicability to related species [J]. Acta Oceanologica Sinica, 200, 29(6): 83 9 [] Sun G H, Yang J M, Song Z L, et al. Isolation of microsatellite markers from Apostichopus japonicus ESTs [J]. Oceanologia et Limnologia Sinica, 200, 4(): [,,,. (Apostichopus japonicus) EST., 200, 4(): 33 38] [2] Chen S L, Liu Y G, Xu M Y, et al. Isolation and characterization of polymorphic microsatellite loci from an EST library of red sea bream (Chrysophrys major) and cross species amplification [J]. Molecular Ecology Notes, 2005, 5(2): [3] Zhan A B, Bao Z M, Wang X L, et al. Microsatellite markers derived from bay scallop Argopecten irradians expressed sequence tag [J]. Fish Science, 2005, 7(6): [4] Yu H, Li Q. Exploiting EST databases for the development and characterization of EST-SSRs in the pacific oyster (Crassostrea gigas) [J]. Journal of Heredity, 2008, 99(2): [5] Wang H X, Li F H, Xiang J H. Polymorphic EST-SSR markers and their mode of inheritance in Fenneropenoeus chinensis [J]. Aquaculture, 2005, 249(4082): 07 4 [6] Li Y F, Liu W D, Gao X G, et al. Construction of cdna libraries from mantle and kidney of Japanese scallop (Mizuhopecten yessoensis) and ESTs analysis [J]. Journal of Fishery Sciences Of China, 200, 7(3): [,,,. cdna EST., 200, 7(3): ] [7] Zhan A B, Bao Z M, Hu X L. Characterization of 95 novel microsatellite markers for Zhikong scallop Chlanys farreri using FIASCO-colony hybridization and EST database mining [J]. Fish Science, 2008, 74(3): [8] Gerber H P, Seipel K, Georgiev O, et al. Transcriptional activation modulated by homopolymeric glutamine and proline stretches [J]. Science, 994, 263(548): [9] Perutz M F, Johnson T, Suzuki M, et al. Glutamine repeats as polar zippers: Their possible role in inherited neurodegenerative diseases [J]. Proceedings of the National Academy of Sciences, 994, 9(2): [20] Gábor T, Zoltán G, Jerzy J. Microsatellites in different eukaryotic genomes: survey and analysis [J]. Genome Research, 2000, 0(7): [2] Schlötterer C. Evolutionary dynamics of microsatellite DNA [J]. Chromosoma, 2000, 09(8): [22] Schlotterer C, Tautz D. Slippage synthesis of simple sequence

Transcriptome analysis and SSR/SNP markers information of the blunt snout bream (Megalobrama amblycephala) [J]. PLoS One, 202, 8(2): e42637 [4] Yuan Z Y, Zhang L L, Zhang G F.

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Microsoft Word - 2-曾聪（new）.doc

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Microsoft Word 曾 聪_new_.doc

Microsoft Word 曾聪_new_.doc