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本研究借助已经构建的本地化EST数据库,利用不同物种对应基因序列进行比对、拼接,成功从大豆中克隆了4个高丝氨酸代谢途径相关酶基因,同时应用已构建的大豆整合图谱,克隆了大豆高丝氨酸代谢途径中7个相关酶基因,并进行了定位分析。通过对氨基酸序列进行比对分析,表明大豆高丝氨酸代谢途径相关酶基因在植物中具有较高的同源性,且序列同源性多大于60%,且与双子叶植物对应序列同源性较高,但与单子叶植物对应基因同源性稍低。从基因定位分析的结果可以看出,7个大豆高丝氨酸代谢途径相关酶基因被定位在D1a、N、B2、G、J和D2六个连锁群上,并同时获得了对应连锁群区间两侧的标记。基因结构分析表明,7个基因的g DNA片段长度介于1 083~5 818 bp之间;c DNA片段长度介于1 083~3 166 bp之间;内含子数目介于0~11个之间,外显子数目介于1~12个之间。本研究为其他氨基酸合成相关酶基因的克隆提供了参考依据,并为基因功能研究以及分子辅助育种打下良好的基础。
In this study, four ESTs related to homoserine metabolism pathway were successfully cloned from soybean by using the constructed EST databases. The corresponding gene sequences of different species were aligned and spliced. The constructed soybean integrated map was successfully cloned Soybean homoserine metabolic pathway in seven related enzyme genes, and the positioning analysis. Comparison of amino acid sequences showed that the homologous genes of soybean homoserine metabolic pathway related genes were higher in plants than those in dicotyledonous plants High, but with monocotyledons corresponding gene homology is slightly lower. According to the results of gene mapping analysis, the seven enzymes related to homoserine metabolism in soybean were mapped on six linkage groups D1a, N, B2, G, J and D2, and the corresponding linkage group interval The mark. Genetic analysis showed that the lengths of g DNA fragments ranged from 1 083 to 5 818 bp, the length of c DNA fragments ranged from 1 083 to 3 166 bp, and the number of introns was from 0 to 11 Between, the number of exons is between 1 to 12. This study provides a reference for the cloning of other amino acid synthesis related enzyme genes, and lay a good foundation for gene function research and molecular assisted breeding.