应用cre/lox植物表达载体的AtNHX1基因转化杨树研究

[目的]研究Na^＋／H^＋逆向转运蛋白AtNHXl基因对杨树的转化。[方法]以杨树南林895杨的叶片作为外植体，采用根癌农杆菌介导法，构建cre／lox植物表达载体，并对3株转基因植株进行PCR分子检测。[结果]结果表明，较适合的转化条件为：外植体在分化培养基上经过2d预培养后于0D600nm值为0．3～0．4的菌液中浸泡7min，卡那抗性绿苗率可达43．9％；PCR分子检测表明，目的基因已经整合到杨树基因组中。[结论]该系统可以应用于杨树的基因工程研究。

Marker_Free: a Novel Tendency of Transgenic Plants

Marker free is a rapidly developed strategy that offers a new approach for the elimination of public concerns caused by the selectable marker genes conferring antibiotic or herbicide resistance and so on. Furthermore, marker_free transgenic plants (MFTPs) have a number of special advantages, such as decreasing the concerns about safety of selectable marker and stacking transgenes progressively into transgenic plants, which significantly owns potential application value. Major approaches developed recently for obtaining MFTPs were reviewed in this paper.

选择标记可去除的植物高效表达载体的构建

在常用的植物组成型表达载体pBI121的选择标记基因NPTII两侧插入同向的lox位点并用多克隆位点(MCS)取代了GUS基因序列,构建了NPTII基因可被去除的和可插入目的基因的通用植物表达载体pBI121-lox-MCS。替换pBI121-lox-MCS中驱动目的基因表达的35S启动子,可构建成一系列具有其他表达特性的植物表达载体,如本文描述的韧皮部特异表达载体pBdENP-lox-MCS。为方便地筛选去除选择标记基因的转基因植物,还构建了绿色荧光蛋白(GFP)表达框与NPTII表达框连锁的pBI121-gfp-lox-MCS载体。上述植物表达载体可广泛应用于培育选择标记可去除的转基因植物。