Lichtenzveig J, Scheuring C, Dodge J, Abbo S, Zhang H-B
Institute of Plant Science and Genetics in Agriculture, The Hebrew University of Jerusalem, Rehovot, 76100, Israel.
Theor Appl Genet. 2005 Feb;110(3):492-510. doi: 10.1007/s00122-004-1857-8. Epub 2004 Dec 11.
Large-insert bacterial artificial chromosome (BAC) libraries, plant-transformation-competent binary BAC (BIBAC) libraries, and simple sequence repeat (SSR) markers are essential for many aspects of genomics research. We constructed a BAC library and a BIBAC library from the nuclear DNA of chickpea, Cicer arietinum L., cv. Hadas, partially digested with HindIII and BamHI, respectively. The BAC library has 14,976 clones, with an average insert size of 121 kb, and the BIBAC library consists of 23,040 clones, with an average insert size of 145 kb. The combined libraries collectively cover ca. 7.0 x genomes of chickpea. We screened the BAC library with eight synthetic SSR oligos, (GA)10, (GAA)7, (AT)10, (TAA)7, (TGA)7, (CA)10, (CAA)7, and (CCA)7. Positive BACs were selected, subcloned, and sequenced for SSR marker development. Two hundred and thirty-three new chickpea SSR markers were developed and characterized by PCR, using chickpea DNA as template. These results have demonstrated that BACs are an excellent source for SSR marker development in chickpea. We also estimated the distribution of the SSR loci in the chickpea genome. The SSR motifs (TAA)n and (GA)n were much more abundant than the others, and the distribution of the SSR loci appeared non-random. The BAC and BIBAC libraries and new SSR markers will provide valuable resources for chickpea genomics research and breeding (the libraries and their filters are available to the public at http://hbz.tamu.edu).
大插入细菌人工染色体（BAC）文库、具备植物转化能力的二元BAC（BIBAC）文库以及简单序列重复（SSR）标记对于基因组学研究的诸多方面至关重要。我们分别用 HindIII 和 BamHI 对鹰嘴豆（Cicer arietinum L.，品种 Hadas）的核 DNA 进行部分酶切，构建了一个 BAC 文库和一个 BIBAC 文库。BAC 文库有 14976 个克隆，平均插入片段大小为 121 kb，BIBAC 文库由 23040 个克隆组成，平均插入片段大小为 145 kb。这两个文库合起来共覆盖了约 7.0 倍鹰嘴豆基因组。我们用八个合成的 SSR 寡核苷酸（GA）10、（GAA）7、（AT）10、（TAA）7、（TGA）7、（CA）10、（CAA）7 和（CCA）7 筛选 BAC 文库。挑选出阳性 BAC，进行亚克隆并测序以开发 SSR 标记。以鹰嘴豆 DNA 为模板，通过 PCR 开发并鉴定了 233 个新的鹰嘴豆 SSR 标记。这些结果表明，BAC 是鹰嘴豆 SSR 标记开发的优良来源。我们还估计了 SSR 位点在鹰嘴豆基因组中的分布。SSR 基序（TAA）n 和（GA）n 比其他基序丰富得多，且 SSR 位点的分布似乎是非随机的。BAC 和 BIBAC 文库以及新的 SSR 标记将为鹰嘴豆基因组学研究和育种提供有价值的资源（这些文库及其滤膜可在 http://hbz.tamu.edu 向公众提供）。
