37 5 Vol. 37, No.5 203 9 ACTA HYDROBIOLOGICA SINICA Sep., 2 0 3 doi: 0.754/203.29 454 GS FLX ESTs 曾 聪, 2 高 泽 霞 罗 伟 刘 肖 莲 王 卫 民 (.,,, 430070; 2., 430070) 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 430070, China; 2. Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan 430070, China) : ; EST-SSR; Key words: Megalobrama amblycephala; EST-SSR; Characteristics 中 图 分 类 号 : Q346 +.5 文 献 标 识 码 : A 文 章 编 号 : 000-3207(203)05-0982-07 (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 (http://www.ncbi. nlm.nih.gov) 60, EST-SSR 收 稿 日 期 : 202-05-30; 修 订 日 期 : 203-02-7 基 金 项 目 : (CARS-46-05); (320988) ( 20PY023 203PY066); 作 者 简 介 : (987 ),, ; ; E-mail: congzeng@live.cn 通 信 作 者 :,, ; E-mail: wangwm@mail.hzau.edu.cn
5 : 454 GS FLX ESTs 983 454 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 (http://code.google.com/p/msatcommander/downloads/list), unigene SSR, SSR SSR 2, 2 SSR 50 bp,, EST 24 36 SSR, 50 bp 2 2. 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 3.93 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, 23.27 ( ),,, 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
984 37
5 : 454 GS FLX ESTs 985 EST EST-SSR 2 32 bp, 2 bp, 6007, ; 5 bp, 343, 322 bp, 6 2 9 bp SSR SSR 84.64%, 20 30 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, 2 4 0 3 67 76, 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,,
986 37 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% 2 3 0 28 25 4 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% 2 3 0 26 8 6 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% 2 4 0 29 39 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% 2 4 0 3 67 76 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], 27 24 bp (69.23%), AC/TG 2 (53.85%),
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