Construction and characterization of a papaya BAC library as a foundation for molecular dissection of a tree-fruit genome
Construction and characterization of a papaya BAC library as a foundation for molecular dissection of a tree-fruit genome
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DOI：
10.1007/s001220000448
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发表时间：
2001-05-01
影响因子：
5.4
通讯作者：
Paterson, AH
中科院分区：
文献类型：
--
作者：
Ming, R;Moore, PH;Paterson, AH
从番木瓜(Carica Papaya L.)幼叶中提取高分子量DNA，构建了细菌人工染色体文库。该文库由来自两个不同连接反应的39168个克隆组成。文库的平均插入片段大小为132kb，第一次连接的18700个克隆中有96.5%的克隆含有平均86kb的插入片段，第二次连接的20468个克隆中有95.7%的克隆含有平均174kb的插入片段。两个高粱叶绿体探针分别与文库杂交，共获得504个叶绿体克隆，占文库的1.4%。整个BAC文库估计提供了13.7倍的木瓜基因组等价物，不包括假阳性克隆和叶绿体克隆。将文库的94%或36864个克隆与12.7x木瓜基因组等效物一起制成高密度过滤器。11个番木瓜和10个拟南芥cDNA探针在文库中平均每个探针检测到22.8个BAC。由于其相对较小的基因组(372MBP/1C)以及在种植后9至15个月就能产生成熟果实的能力，木瓜显示出作为研究影响结果性状的基因的模式植物的前景。一种快速定位水果控制基因的方法将是根据EST与之杂交的BAC重叠群组装物理图谱。木瓜基因组的物理图谱将大大增强我们克隆和操纵具有经济重要性的基因的能力。
A bacterial artificial chromosome (BAC) library was constructed from high-molecular-weight DNA isolated from young leaves of papaya (Carica papaya L.). This BAC library consists of 39168 clones from two separate ligation reactions. The average insert size of the library is 132 kb; 96.5% of the 18700 clones from the first ligation contained inserts that averaged 86 kb in size, 95.7% of the 20468 clones from the second Ligation contained inserts that averaged 174 kb in size. Two sorghum chloroplast probes hybridized separately to the library and revealed a total of 504 chloroplast clones or 1.4% of the library. The entire BAC library was estimated to provide 13.7 x papaya-genome equivalents, excluding the false-positive and chloroplast clones. High-density filters were made containing 94% or 36864 clones of the library with 12.7 x papaya-genome equivalents. Eleven papaya-cDNA and ten Arabidopsis-cDNA probes detected an average of 22.8 BACs per probe in the library. Because of its relatively small genome (372 Mbp/1 C) and its ability to produce ripe fruit 9 to 15 months after planting, papaya shows promise as a model plant studying genes that affect fruiting characters. A rapid approach to locating fruit-controlling genes will be to assemble a physical map based on BAC contigs to which ESTs have hybridized. A physical map of the papaya genome will significantly enhance our capacity to clone and manipulate genes of economic importance.