Soybean [Glycine max (L.) Merr.] embryogenic cultures were transformed by particle bombardment with the feedback-insensitive tobacco anthranilate synthase (AS) gene ASA2 driven by the CaMV 35S promoter and selected using hph as the selectable marker gene. Only one of eight regenerated lines that set seed and contained ASA2 expressed the gene highly and contained increased free tryptophan (Trp) levels in leaves, seeds and embryogenic cultures. Leaf extracts of the ASA2 expressing line contained about twice as much AS enzyme activity as the untransformed control and this activity was only slightly more feedback-insensitive. Amino acid analysis showed that both leaves and embryogenic tissue cultures of the ASA2 expressing line had four to five-times the normal levels of free Trp and slightly higher free tyrosine and phenylalanine. The seed total Trp content was only slightly increased. Metabolic profiling-analysis by GC-MS detected no other consistent differences. These studies show that the ASA2 gene can be expressed in soybean and that modest changes in Trp synthesis occurs.

译文

:大豆[Glycine max(L.)Merr。]胚性培养物通过由CaMV 35S启动子驱动的反馈不敏感的烟草邻氨基苯甲酸合酶(AS)基因ASA2进行粒子轰击进行转化,并使用hph作为选择标记基因进行选择。八个定植种子并含有ASA2的再生系中只有一个表达该基因,并且在叶片,种子和胚发生培养物中的游离色氨酸(Trp)水平升高。表达ASA2的品系的叶提取物所含的AS酶活性约为未转化对照的两倍,该活性对反馈的敏感性稍高。氨基酸分析表明,ASA2表达株系的叶片和胚发生组织培养物的游离Trp含量是正常水平的4至5倍,游离酪氨酸和苯丙氨酸含量稍高。种子总色氨酸含量仅略有增加。 GC-MS进行的代谢谱分析没有发现其他一致的差异。这些研究表明,ASA2基因可以在大豆中表达,并且Trp合成发生适度的变化。

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