金鱼hAT家族转座子Tgf2的克隆及其结构

doi:10.3724/SP.J.1005.2010.01263

遗传 ›› 2010, Vol. 32 ›› Issue (12): 1263-1268.doi: 10.3724/SP.J.1005.2010.01263

金鱼hAT家族转座子Tgf2的克隆及其结构

邹曙明, 杜雪地, 袁剑, 蒋霞云

上海海洋大学农业部种质资源与利用重点开放实验室, 上海201306

收稿日期:2010-03-23 修回日期:2010-05-08 出版日期:2010-12-20 发布日期:2010-12-20
通讯作者: 邹曙明 E-mail:smzou@shou.edu.cn
基金资助:
国家高技术研究发展计划(863计划)项目(编号：2009AA10Z105)、公益性行业科研专项(编号：200903045)和上海市教委曙光计划 (编号：08SG50)资助

Cloning of goldfish hAT transposon Tgf2 and its structure

ZOU Shu-Ming, DU Xue-Di, YUAN Jian, JIANG Xia-Yun

Key Laboratory of Aquatic Genetic Resources and Utilization, Ministry of Agriculture, Shanghai Ocean University, Shanghai 201306, China

Received:2010-03-23 Revised:2010-05-08 Online:2010-12-20 Published:2010-12-20
Contact: ZOU Shu-Ming E-mail:smzou@shou.edu.cn

摘要/Abstract

摘要： hAT家族转座子以果蝇hobo、玉米Ac和金鱼草(Ceratophyllum demersum L.) Tam3为代表, 以“剪切－粘帖”方式进行DNA转座。1996年, 日本学者首次在白化青鳉(Oryzias latipes)中发现具有天然活性的脊椎动物hAT家族转座子, 即青鳉Tol2转座子, 该转座子已在模式生物斑马鱼转基因、基因和启动子捕获方面进行了广泛应用。文章根据玉米Ac与青鳉Tol2转座子序列保守区设计一对引物, 在19种不同鱼类物种或品系中进行PCR筛选, 最后发现此类hAT家族转座子在我国不同品系金鱼中存在, 命名为金鱼Tgf2转座子。金鱼Tgf2转座子全长4 720 bp, 由4个阅读框组成, 与青鳉Tol2转座子的相似度为97%。金鱼Tgf2与青鳉Tol2转座子在末端倒位重复和亚末端重复上存在一定差异, 此外, 金鱼Tgf2转座子的中间反向重复序列(1 453 bp到2 091 bp)可形成一种“十”字结构, 明显有别于青鳉Tol2转座子形成的茎环结构, 这些区域与转座活性密切相关。文章预示金鱼Tgf2转座子可能具有更高的天然转座活性, 构建高效金鱼Tgf2转基因元件可供鱼类转基因和基因捕获研究。

关键词: 金鱼, hAT家族, Tgf2转座子, 转基因, 基因捕获

Abstract: The hAT transposon family, including hobo of Drosophila, Ac of maize (Zea mays L.) and Tam3 of snapdragon (Ceratophyllum demersum L.), is proposed to be involved in transposition between genomic DNAs in "cut and paste" pat-terns. In 1996, a transposon of Tol2, the first autonomous transposon in vertebrate, was identified from the genome of albino medaka fish (Oryzias latipes). Since then, a new transgenic and gene trap system based on Tol2 has been developed and widely used in zebrafish. In this study, we designed gene-specific primers based on the conserved regions of amino acid sequences between medaka Tol2 and maize Ac. Meanwhile, PCR was carried out in 19 fish species or strains including goldfish. Finally, another similar hAT transposon, termed as Tgf2, was identified in the genomes from different strains of goldfish. Goldfish Tgf2 is 4,720 bp in length including 4 exons and shares an identity of 97% with medaka Tol2. Distinct differences were observed in the terminal inverted repeat (TIR) and subterminal repeat (STR) regions between Tgf2 and Tol2. In addition, the internal inverted repeat (IIR) region of Tgf2 (1453 bp-2091 bp) tended to form a "+" crossing structure, rather than a stem-loop structure in medaka Tol2. The regions, including TIR, STR, and IIR, were supposed to be closely related to the transposing activities of hAT transposon family members. From these sequence differences, we may expect a probably high activity of goldfish Tgf2 in transposition and it could be further used as a tool for transgenesis and gene trap in aquaculture fish in future.

Key words: Tgf2, goldfish, Carassius auratus, transposon

邹曙明，杜雪地，袁剑，蒋霞云. 金鱼hAT家族转座子Tgf2的克隆及其结构[J]. 遗传, 2010, 32(12): 1263-1268.

JU Shu-Meng, DU Xue-De, YUAN Jian, JIANG Xia-Yun. Cloning of goldfish hAT transposon Tgf2 and its structure[J]. HEREDITAS, 2010, 32(12): 1263-1268.

参考文献

[1] McClintock B. Chromosome organization and genic ex-pression. Cold Spring Harbor Symp Quant Biol, 1951, 16(3): 13–17. [2] Murai N, Li ZJ, Kawagoe Y, Hayashimoto A. Transposi-tion of the maize activator element in transgenetic rice plants. Nucleic Acids Res, 1991, 19(3): 617–622. [3] Amutan M, Nyyssönen E, Stubbs J, Diaz-Torres MR, Dunn-Coleman N. Identification and cloning of a mobile transposon from Aspergillus niger var. awamori. Curr Genet, 1996, 29(5): 468–473. [4] 谢飞, 高波, 宋成义, 陈国宏. “睡美人”转座子的研究进展. 遗传, 2007, 29(7): 785–792. [5] Koga A, Suzuki M, Inagaki H, Bessho Y, Hori H. Trans-posable element in fish. Nature, 1996, 383: 30. [6] Kawakami K, Shima A. Identification of the Tol2 transpo-sase of the medaka fish Oryzias latipes that catalyzes excision of a nonautonomous Tol2 element in zebrafish Danio rerio. Gene, 1999, 240(1): 239–244. [7] Urasaki A, Mito T, Noji S, Ueda R, Kawakami K. Trans-position of the vertebrate Tol2 transposable element in Drosophila melanogaster. Gene, 2008, 425(1–2): 64–68. [8] Kawakami K, Koga A, Hori H, Shima A. Excision of the Tol2 transposable element of the medaka fish, Oryzias latipes, in zebrafish, Danio rerio. Gene, 1998, 225(1–2):17–22. [9] Kawakami K, Shima A, Kawakami N. Identification of a functional transposase of the Tol2 element, an Ac-like element from the Japanese medaka fish, and its transposi-tion in the zebrafish germ lineage. Proc Nalt Acad Sci USA, 2000, 97(21): 11403–11408. [10] Kawakami K, Imanaka K, Itoh M, Taira M. Excision of the Tol2 transposable element of the medaka fish Oryzias latipes in Xenopus laevis and Xenopus tropicalis. Gene, 2004, 338(1): 93–98. [11] Sato Y, Kasai T, Nakagawa S, Tanabe K, Watanabe T, Kawakami K, Takahashi Y. Stable integration and condi-tional expression of electroporated transgenes in chicken embryos. Dev Biol, 2007, 305(2): 616–624. [12] Kawakami K, Noda T. Transposition of the Tol2 element, an Ac-like element from the Japanese medaka fish Oryzias latipes, in mouse embryonic stem cells. Genetics, 2004, 166(2): 895–899. [13] Balciunas D, Wangensteen KJ, Wilber A, Bell J, Geurts A, Sivasubbu S, Wang X, Hackett PB, Largaespada DA, McIvor RS, Ekker SC. Harnessing a high cargo-capacity transposon for genetic applications in vertebrates. PLoS Genet, 2006, 2(11): e169. [14] Wu SCY, Meir YJJ, Coates CJ, Handler AM, Pelczar P, Moisyadi S, Kaminski JM. PiggyBac is a flexible and highly active transposon as compared to sleeping beauty, Tol2, and Mos1 in mammalian cells. Proc Natlt Acad Sci USA, 2006, 103(41): 15008–15013. [15] Koichi Kawakami K, Takeda H, Kawakami N, Kobayashi M, Matsuda N, Mishina M. A transposon-mediated gene trap approach identifies developmentally regulated genes in zebrafish. Dev Cell, 2004, 7(1): 133–144. [16] Kotani T, Nagayoshi S, Urasaki A, Kawakami K. Trans-poson-mediated gene trapping in zebrafish. Methods, 2006, 39(3): 199–206. [17] Fisher S, Grice EA, Vinton RM, Bessling SL, Urasaki A, Kawakami K, McCallion AS. Evaluating the biological relevance of putative enhancers using Tol2 transpo-son-mediated transgenesis in zebrafish. Nat Protoc, 2006, 1(3): 1297–1305. [18] Asakawa K, Suster ML, Mizusawa K, Nagayoshi S, Kotani T, Urasaki A, Kishimoto Y, Hibi M, and Kawakami K. Genetic dissection of neural circuits by Tol2 transpo-son-mediated Gal4 gene and enhancer trapping in zebraf-ish. Proc Nalt Acad Sci USA, 2008, 105(4): 1255–1260. [19] Nagayoshi S, Hayashi E, Abe G, Osato N, Asakawa K, Urasaki A, Horikawa K, Ikeo K, Takeda H, Kawakami K. Insertional mutagenesis by the Tol2 transposon-mediated enhancer trap approach generated mutations in two de-velopmental genes: tcf7 and synembryn-like. Development, 2008, 135(1): 159–169. [20] Parinov S, Kondrichin I, Korzh V, Emelyanov A. Tol2 transposon-mediated enhancer trap to identify develop-mentally regulated zebrafish genes in vivo. Dev Dyn, 2004, 231(2): 449–459. [21] Kawakami K. Tol2: a versatile gene transfer vector in ver-tebrates. Genome Biol, 2007, 8(Suppl. 1): S7. [22] Urasaki A, Morvan G, Kawakami K. Functional dissection of the Tol2 transposable element identified the minimal cis-sequence and a highly repetitive sequence in the sub-terminal region essential for transposition. Genetics, 2006, 174(2): 639–649. [23] Sambrook J, Fritsch EF, Maniatis T. Molecular Cloning: A Laboratory Manual. 2nd ed. Cold Spring Harbor Labora-tory, Cold Spring Harbor, NY, 1989. [24] Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res, l994, 22(22): 4673–4680. [25] Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecu-lar evolutionary genetics analysis (MEGA) software ver-sion 4.0. Mol Biol Evol, 2007, 24(8): 1596–1599. [26] 付光明, 苏乔, 安利佳. 青鳉Tol2转座子研究进展. 遗传, 2006, 28(7): 899–905. [27] Koga A, Hori H. The Tol2 transposable element of the medaka fish: an active DNA-based element naturally oc-curring in a vertebrate genome. Genes Genet Syst, 2001, 76(1): 1–8. [28] 王鸿媛, 王得良, 张词祖, 张斌. 金鱼. 北京: 中国林业出版社, 2003, 1–5. [29] Koga A, Shimada A, Shima A, Sakaizumi M, Tachida H, Hori H. Evidence for recent invasion of the medaka fish genome by the Tol2 transposable element. Genetics, 2000, 155(1): 273–281.

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金鱼hAT家族转座子Tgf2的克隆及其结构

Cloning of goldfish hAT transposon Tgf2 and its structure

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