Frohme M, Camargo A A, Heber S, Czink C, Simpson A J, Hoheisel J D, de Souza A P
Functional Genome Analysis, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 506, D-69120 Heidelberg, Germany.
Nucleic Acids Res. 2000 Aug 15;28(16):3100-4. doi: 10.1093/nar/28.16.3100.
A cosmid library was made of the 2.7 Mb genome of the Gram-negative plant pathogenic bacterium Xylella fastidiosa and analysed by hybridisation mapping. Clones taken from the library as well as genomic restriction fragments of rarely cutting enzymes were used as probes. The latter served as a backbone for ordering the initial map contigs and thus facilitated gap closure. Also, the co-linearity of the cosmid map, and thus the eventual sequence, could be confirmed by this process. A subset of the eventual clone coverage was distributed to the Brazilian X.FASTIDIOSA: sequencing network. Data from this effort confirmed more quantitatively initial results from the hybridisation mapping that the redundancy of clone coverage ranged between 0 and 45-fold across the genome, while the average was 15-fold by experimental design. Reasons for this not unexpected fluctuation and the actual gaps are being discussed, as is the use of this effect for functional studies.
构建了革兰氏阴性植物病原菌木质部难养菌2.7 Mb基因组的黏粒文库,并通过杂交作图进行分析。从文库中选取的克隆以及稀有切割酶的基因组限制片段用作探针。后者作为排序初始图谱重叠群的主干,从而便于填补缺口。此外,通过该过程可以确认黏粒图谱的共线性,进而确认最终序列的共线性。最终克隆覆盖的一个子集被分配到巴西木质部难养菌测序网络。这项工作的数据更定量地证实了杂交作图的初步结果,即整个基因组中克隆覆盖的冗余度在0至45倍之间,而根据实验设计平均为15倍。正在讨论这种意料之中的波动以及实际缺口的原因,以及这种效应在功能研究中的应用。