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首页> 外文期刊>Acta Horticulturae >BAG Sequencing and Annotation Driven by Experimental Data: an Application for a Serine-threonine Kinase on Tomato Chromosome 12
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BAG Sequencing and Annotation Driven by Experimental Data: an Application for a Serine-threonine Kinase on Tomato Chromosome 12

机译：实验数据驱动的BAG测序和注释：丝氨酸苏氨酸激酶在番茄12号染色体上的应用

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摘要

Through the International Tomato Sequencing Project, the first BAG (Bacterial Artificial Chromosome) of chromosome 12 (Le_HBa0032K07) has been sequenced and submitted to the Solanaceae Genomics Network (SGN) repository. The preliminary annotation of the BAC was based on comparisons with a collection of about 215,000 Expressed Sequence Tags (ESTs) from tomato species and on their tentative consensus sequences (TCs). Considering the ESTs/TCs-to-genome alignments, 11 gene loci were located on this BAC.One of these was detected at the SP6 end of the BAC where a partial match of a tentative consensus sequence, generated by assembling 24 ESTs, was found. In order to verify both the reliability of the computationally detected gene locus and of its expressed product, we designed primers on the complete sequence of the TC. Then polymerase chain reactions (PCRs) were carried out: a 5000 bp fragment was amplified on genomic DNA, while a 500 bp fragment was obtained from cDNA amplification, as expected. Thisfragment was sequenced and then compared to the TC. The result of the sequence comparison confirms both the reliability of the TC and the expression of the gene. Finally, using the SGN BAC-ends database, we selected four BACs in order to extend the seedBAC Le_HBa0032K07 and to cover the full length of the TC.
机译：通过国际番茄测序项目，已对12号染色体的第一个BAG（细菌人工染色体）（Le_HBa0032K07）进行了测序，并提交给茄科基因组网络（SGN）信息库。 BAC的初步注释是基于与来自番茄物种的约215,000个表达序列标签（EST）及其暂定共有序列（TC）的比较。考虑到ESTs / TCs与基因组的比对，在该BAC上定位了11个基因位点，其中一个在BAC的SP6末端检测到，其中发现了通过组装24个ESTs而产生的暂时性共有序列的部分匹配。 。为了验证计算检测到的基因座及其表达产物的可靠性，我们在TC的完整序列上设计了引物。然后进行聚合酶链反应（PCR）：按照预期，在基因组DNA上扩增了5000 bp的片段，而从cDNA扩增获得了500 bp的片段。对该片段进行测序，然后与TC进行比较。序列比较的结果证实了TC的可靠性和基因的表达。最后，使用SGN BAC-ends数据库，我们选择了四个BAC，以扩展seedBAC Le_HBa0032K07并覆盖TC的整个长度。

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