大豆高丝氨酸代谢途径相关酶基因的电子克隆与定位分析

In Silico Cloning and Mapping Analysis of Genes Related to Homoserine Metabolism in Soybean

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 thi s research, four genes related to homoserine metabolism in soybean were cloned by using the local EST database with comparison and blasting the gene sequences from different species. As well as seven genes involved in the homoserine metabolism in soybean were cloned and mapped. Comparisons analysis of amino acid sequences showed that the genes related to homoserine metabolism in soybean exhibited high sequence similarity（over than 60%） with other homologs in plants. Moreover, the sequences displayed higher similarity in dicotyledons than in monocotyledons. Gene mapping analysis indicated that the seven enzyme genes were further mapped into the six linkages groups of soybean genome, including D1 a, N, B2, G, J and D2, and the flanking markers of these genes on the linkage group were obtained respectively. The analysis of gene structure showed that the g DNA length of these seven genes were 1 083-5 818 bp, and the c DNA length of these seven genes were 1 083-3 166 bp with 0 to 11 introns, and 1 to 12 exons in these genes. These results provided areference for further cloning of the genes related to other amino acid synthesis, and also provided a foundation for the functional research of genes in molecular assisted selection.