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EGTC; Database for the Exchangeable Gene Trap Clones

机译：EGTC；可交换基因陷阱克隆的数据库

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Gene trapping in ES cells is a proven method for large-scale random insertional mutagenesis in the mouse genome. Gene trap vectors contain a promoter-less reporter gene downstream of a splice acceptor and a selectable marker gene. A fusion transcript between the integrated (trapped) gene and the reporter gene can be easily cloned by 5'-RACE. We have developed exchangeable gene trap vectors, pU18, pU21, pU21B, pU21T and pU22. The reporter beta-geo gene can be exchanged into any other DNA of interest through Cre-mediated recombination. We have isolated trap ES clones, determined sequence tags (GSSs) of trapped genes by 5'-RACE, and annotated them by BLAST and BLAT search. The data of trap clones are opened as a Database for the Exchangeable Gene Trap Clones, EGTC. Until the middle of JAN. 2008, total 406 GSSs have been registered. Among them, 75% are known genes, 16% are ESTs, and 9% are new genes. 332 GSSs were determined their chromosomal localization through BLAT search. Since the pU21, pU21B and pU21T vectors have stop codons in upstream of the start codon of the beta-geo, the integration sites of the vectors showed a strong bias to the vicinity of the ATG exon of trapped genes, indicating that these vectors could induce a null allele efficiently. In addition to determination of GSSs, 141 trap mouse lines have been established and deposited to the CARD R-BASE; the database for cryopreserved embryos.
机译：ES细胞中的基因捕获是在小鼠基因组中大规模随机插入诱变的一种行之有效的方法。基因捕获载体在剪接受体下游包含无启动子的报告基因和选择标记基因。整合的（捕获的）基因和报告基因之间的融合转录本可以很容易地通过5'-RACE克隆。我们已经开发了可互换的基因陷阱载体，pU18，pU21，pU21B，pU21T和pU22。可以通过Cre介导的重组将报告基因beta-geo基因交换成任何其他感兴趣的DNA。我们已分离出陷阱ES克隆，通过5'-RACE确定了被捕获基因的序列标签（GSS），并通过BLAST和BLAT搜索对其进行了注释。陷阱克隆的数据作为可交换基因陷阱克隆EGTC的数据库打开。直到1月中旬。 2008年，总共注册了406个GSS。其中，已知基因为75％，EST为16％，新基因为9％。通过BLAT搜索确定了332个GSS的染色体定位。由于pU21，pU21B和pU21T载体在β-geo起始密码子的上游具有终止密码子，因此这些载体的整合位点对捕获基因的ATG外显子附近表现出强烈的偏见，表明这些载体可以诱导无效等位基因。除了确定GSS外，还建立了141个陷阱小鼠系并将其保存到CARD R-BASE中。冷冻保存的胚胎数据库。

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