Sahab Sareena, Taylor Nigel
Agriculture Victoria Research, AgriBio, Centre for AgriBioscience, Bundoora, VIC, Australia.
Donald Danforth Plant Science Center, St. Louis, MO, United States.
Front Plant Sci. 2022 Feb 4;13:816323. doi: 10.3389/fpls.2022.816323. eCollection 2022.
Imperfect T-DNA processing is common during -mediated transformation, which integrates vector backbone sequences into the plant genome. However, regulatory restrictions prevent such transgenic plants from being developed for commercial deployment. The binary vector pCAMBIA2300 was modified by incorporating multiple left border (Mlb) repeats and was tested in BY2 cells, tobacco, and cassava plants to address this issue. PCR analyses confirmed a twofold increase in the vector backbone free events in the presence of triple left borders in all three systems tested. Vector backbone read-through past the LB was reduced significantly; however, the inclusion of Mlbs did not effectively address the beyond right border read-through. Also, Mlbs increased the frequency of single-copy and vector backbone free events (clean events) twice compared to a single LB construct. Here, we briefly narrate the strength and limitations of using Mlb technology and reporter genes in reducing the vector backbone transfer in transgenic events.
在农杆菌介导的转化过程中，不完美的T-DNA加工很常见，这会将载体骨架序列整合到植物基因组中。然而，监管限制使得此类转基因植物无法用于商业种植。二元载体pCAMBIA2300通过整合多个左边界（Mlb）重复序列进行了修饰，并在BY2细胞、烟草和木薯植株中进行了测试，以解决这一问题。PCR分析证实，在所有测试的三个系统中，存在三重左边界时，无载体骨架事件增加了两倍。载体骨架越过左边界的通读显著减少；然而，包含Mlb并不能有效解决越过右边界的通读问题。此外，与单个LB构建体相比，Mlb使单拷贝和无载体骨架事件（纯合事件）的频率增加了两倍。在此，我们简要叙述了使用Mlb技术和报告基因在减少转基因事件中载体骨架转移方面的优势和局限性。
