Huang Yao-Xiong, Li Lin, Yang Liu, Zhang Yi
Department of Biomedical Engineering, Ji Nan University, Guang Zhou, China.
Biomed Opt Express. 2018 Mar 21;9(4):1783-1794. doi: 10.1364/BOE.9.001783. eCollection 2018 Apr 1.
Here we report a technique of laser chromosome welding that uses a violet pulse laser micro-beam for welding. The technique can integrate any size of a desired chromosome fragment into recipient chromosomes by combining with other techniques of laser chromosome manipulation such as chromosome cutting, moving, and stretching. We demonstrated that our method could perform chromosomal modifications with high precision, speed and ease of use in the absence of restriction enzymes, DNA ligases and DNA polymerases. Unlike the conventional methods such as de novo artificial chromosome synthesis, our method has no limitation on the size of the inserted chromosome fragment. The inserted DNA size can be precisely defined and the processed chromosome can retain its intrinsic structure and integrity. Therefore, our technique provides a high quality alternative approach to directed genetic recombination, and can be used for chromosomal repair, removal of defects and artificial chromosome creation. The technique may also have applicability on the manipulation and extension of large pieces of synthetic DNA.
在此，我们报告一种激光染色体焊接技术，该技术使用紫光脉冲激光微束进行焊接。通过与激光染色体操作的其他技术（如染色体切割、移动和拉伸）相结合，该技术可将任何大小的所需染色体片段整合到受体染色体中。我们证明，在没有限制酶、DNA连接酶和DNA聚合酶的情况下，我们的方法能够以高精度、高速度且易于使用的方式进行染色体修饰。与诸如从头人工染色体合成等传统方法不同，我们的方法对插入的染色体片段大小没有限制。插入的DNA大小可以精确界定，且处理后的染色体能够保持其固有结构和完整性。因此，我们的技术为定向基因重组提供了一种高质量的替代方法，可用于染色体修复、缺陷去除和人工染色体创建。该技术在大片段合成DNA的操作和扩展方面可能也具有适用性。
