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首页> 外文期刊>Marine genomics >Isolation and characterization of a genomic fosmid clone containing hspb1 (hsp27) from the Pacific bluefin tuna Thunnus orientalis
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Isolation and characterization of a genomic fosmid clone containing hspb1 (hsp27) from the Pacific bluefin tuna Thunnus orientalis

机译：太平洋蓝鳍金枪鱼Thunnus Orientalis含hspb1（hsp27）的基因组化石克隆的分离与鉴定

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摘要

We constructed a fosmid library of genomic DNA from the Pacific bluefin tuna Thunnus orientalis and isolated a clone containing the complete gene hspb1 (hsp27) from the library. To screen the library, we first obtained a full-length hspb1 cDNA by RACE-PCR. This cDNA encodes a putative protein of 202 amino acids. Phylogenetic analysis indicated that the protein belongs to the Hsp27 family. Northern blot analysis demonstrated that the gene transcript was expressed in tuna muscle tissues. Using a combination of PCR screening and colony hybridization, we isolated a 39,422-bp fosmid clone containing hspb1 from an arrayed library. Sequence alignment showed that hspb1 contains three exons and two introns. Comparative genomic analysis revealed that hspb1 lies in a region of conserved synteny in the genomes of fish and human. Our finding of conserved regions within and around hspb1 in fish species will help to identify functional elements such as promoter regions.
机译：我们从太平洋蓝鳍金枪鱼东方金枪鱼构建了一个基因组DNA fosmid文库，并从该文库中分离了一个包含完整基因hspb1（hsp27）的克隆。为了筛选该文库，我们首先通过RACE-PCR获得了全长hspb1 cDNA。该cDNA编码202个氨基酸的推定蛋白质。系统发育分析表明该蛋白属于Hsp27家族。 Northern印迹分析表明该基因转录物在金枪鱼肌肉组织中表达。使用PCR筛选和菌落杂交的组合，我们从一个阵列文库中分离了一个39,422-bp的含有hspb1的fosmid克隆。序列比对显示hspb1包含三个外显子和两个内含子。比较基因组分析表明，hspb1位于鱼类和人类基因组的保守同义区域。我们在鱼类中hspb1内和周围的保守区域的发现将有助于鉴定功能元件，例如启动子区域。

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