Construction and characterization of a BAC library of soybean |
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Authors: | Zhengjun Xia Hiroko Sato Satoshi Watanabe Shiji Kawasaki Kyuya Harada |
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Affiliation: | (1) Faculty of Horticulture, Chiba University, Matsudo, Chiba 271-8510, Japan;(2) National Institute of Agrobiological Resources, Tsukuba Science City, Ibaraki 305, Japan;(3) Present address: National Institute of Livestock and Grassland Science, 768 Senbonmatu, Nishinasuno, Tochigi 329-2793, Japan |
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Abstract: | We have constructed a soybean (Glycine max (L.) Merrill) bacterial artificial chromosome (BAC) library from green leaf protoplasts of the cultivar, Misuzudaizu. The library contains 53,760 clones with an average insert size of 116 kb. About 2.9% chloroplast DNA origin was revealed by PCR and colony hybridization. Apart from 2.8% clones having no insert, this library represents 5.2 genome equivalents. With this genome coverage, the probability of having any DNA sequence represented in the library is higher than 99.5%. Three-dimensional pools of the BAC library in combination with the use of a high efficiency genome scanning (HEGS) electrophoresis facilitate rapid and efficient PCR-based screening. An average of five positive clones were identified after screening the BAC library with SSR and STS markers. BAC-end walking was performed for three SSR associated BACs. This library will provide a good resource for positional cloning of agronomically and biologically important QTL genes that Misuzudaizu possesses. |
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Keywords: | BAC library flowering time QTL soybean SSR |
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