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细菌人工染色体(bacterial artificial chromosome,BAC)文库是开展基因组测序、基因图位克隆、分子标记开发、物理作图等研究的重要基因组资源。本研究以二倍体野生棉雷蒙德氏棉(G.raimondii,D5)为材料,从成株期暗培养的黄化幼嫩叶片中得到纯净、完整和高质量的基因组DNA。经脉冲场凝胶电泳检测,所提取基因组DNA大于700 kb。经酶切、片段选择、连接转化,构建了雷蒙德氏棉的基因组BAC文库,文库共包含26 880个克隆,平均插入片段为127 kb,覆盖该棉种全基因组的3.7倍左右,克隆空载率小于5%。该文库的构建,为雷蒙德氏棉基因组物理图谱的构建、功能基因的定位与克隆、以及比较基因组研究提供了重要的基因组资源。
The bacterial artificial chromosome (BAC) library is an important genomic resource for genome sequencing, gene mapping, molecular marker development and physical mapping. In this study, G.raimondii (D5) was used as a material to obtain pure, complete and high-quality genomic DNA from adult yellowing young leaves of adult plants. After pulsed field gel electrophoresis, the extracted genomic DNA was greater than 700 kb. A total of 26 880 clones were constructed, with the average insert size of 127 kb, covering 3.7 times of the whole genome of cotton cultivars. Clone The loading rate is less than 5%. The construction of this library provided important genomic resources for the construction of physical map of cotton genome, locating and cloning of functional genes, and comparative genomics.