Churcher C, Bowman S, Badcock K, Bankier A, Brown D, Chillingworth T, Connor R, Devlin K, Gentles S, Hamlin N, Harris D, Horsnell T, Hunt S, Jagels K, Jones M, Lye G, Moule S, Odell C, Pearson D, Rajandream M, Rice P, Rowley N, Skelton J, Smith V, Barrell B
The Sanger Centre, Wellcome Trust Genome Campus, Hinxton, Cambridge, UK.
Nature. 1997 May 29;387(6632 Suppl):84-7.
Large-scale systematic sequencing has generally depended on the availability of an ordered library of large-insert bacterial or viral genomic clones for the organism under study. The generation of these large insert libraries, and the location of each clone on a genome map, is a laborious and time-consuming process. In an effort to overcome these problems, several groups have successfully demonstrated the viability of the whole-genome random 'shotgun' method in large-scale sequencing of both viruses and prokaryotes. Here we report the sequence of Saccharomyces cerevisiae chromosome IX, determined in part by a whole-chromosome 'shotgun', and describe the particular difficulties encountered in the random 'shotgun' sequencing of an entire eukaryotic chromosome. Analysis of this sequence shows that chromosome IX contains 221 open reading frames (ORFs), of which approximately 30% have been sequenced previously. This chromosome shows features typical of a small Saccharomyces cerevisiae chromosome.
大规模系统测序通常依赖于为所研究的生物体构建一个有序的大插入片段细菌或病毒基因组克隆文库。构建这些大插入片段文库以及确定每个克隆在基因组图谱上的位置是一个费力且耗时的过程。为了克服这些问题,几个研究小组已成功证明了全基因组随机“鸟枪法”在病毒和原核生物大规模测序中的可行性。在此,我们报告了酿酒酵母IX号染色体的序列,该序列部分是通过全染色体“鸟枪法”测定的,并描述了在对整个真核染色体进行随机“鸟枪法”测序过程中遇到的特殊困难。对该序列的分析表明,IX号染色体包含221个开放阅读框(ORF),其中约30%以前已被测序。这条染色体展现出酿酒酵母小染色体的典型特征。