The Berkeley Drosophila Genome Project gene disruption project: Single P-element insertions mutating 25% of vital Drosophila genes.
The Berkeley Drosophila Genome Project gene disruption project: Single P-element insertions mutating 25% of vital Drosophila genes.
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发表时间：
1999-08
期刊：
影响因子：
3.3
通讯作者：
N. Mozden;H. Misra;E. Rhem;D. Beaton;David I. Stern;T. Laverty;C. Stern;T. Laverty;Rachael D. Rubin;A. Spradling;G. Rubin;A. Spradling;D. Ho
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作者：
N. Mozden;H. Misra;E. Rhem;D. Beaton;David I. Stern;T. Laverty;C. Stern;T. Laverty;Rachael D. Rubin;A. Spradling;G. Rubin;A. Spradling;D. Ho
遗传学和功能基因组学的一个基本目标是识别和突变模式生物中的每一个基因，如果蝇。伯克利果蝇基因组计划（BDGP）基因破坏项目产生单个p元素插入菌株，每个菌株突变独特的基因组开放阅读框。这些菌株有力地促进了对断裂位点的进一步遗传和分子研究，但P元素是否可以用于突变所有果蝇基因仍不清楚。我们现在报告说，主要的收集已经增长到包含1045个菌株，这些菌株破坏了大约3600个果蝇基因中的25%以上，这些基因对成虫的生存能力至关重要。在这些P插入中，67%已被基因检测证实会导致相关的隐性突变表型，其余大多数系的有效性是根据统计学依据预测的。bbbb920插入的侧翼序列已被确定在基因组中准确定位，并从EST序列集合中鉴定出376个可能受影响的转录本。BDGP收集的菌株可从布卢明顿库存中心获得，并已帮助研究界表征了250个果蝇基因。这里报告了收集的131个额外基因的可能身份。我们的研究结果表明，果蝇基因对P元素失活具有广泛的敏感性，并为完全基于插入位点测序的BDGP初级收集提供了基本原理。我们预测，这种方法可以使85%的果蝇开放阅读框处于实验控制之下。
A fundamental goal of genetics and functional genomics is to identify and mutate every gene in model organisms such as Drosophila melanogaster. The Berkeley Drosophila Genome Project (BDGP) gene disruption project generates single P-element insertion strains that each mutate unique genomic open reading frames. Such strains strongly facilitate further genetic and molecular studies of the disrupted loci, but it has remained unclear if P elements can be used to mutate all Drosophila genes. We now report that the primary collection has grown to contain 1045 strains that disrupt more than 25% of the estimated 3600 Drosophila genes that are essential for adult viability. Of these P insertions, 67% have been verified by genetic tests to cause the associated recessive mutant phenotypes, and the validity of most of the remaining lines is predicted on statistical grounds. Sequences flanking >920 insertions have been determined to exactly position them in the genome and to identify 376 potentially affected transcripts from collections of EST sequences. Strains in the BDGP collection are available from the Bloomington Stock Center and have already assisted the research community in characterizing >250 Drosophila genes. The likely identity of 131 additional genes in the collection is reported here. Our results show that Drosophila genes have a wide range of sensitivity to inactivation by P elements, and provide a rationale for greatly expanding the BDGP primary collection based entirely on insertion site sequencing. We predict that this approach can bring >85% of all Drosophila open reading frames under experimental control.