We propose a method to determine the topological charge (TC) of a perfect vortex. With the phase shift technique, the perfect vortex and its conjugate beam exactly overlap and interfere. Consequently, the TC of a perfect vortex is determined by counting the number of interference fringes. This proposed method enables in situ determination of the TC of the perfect vortex without the need for additional optical elements, and it is immune to environmental vibration and parasitic interference.
Unlike eukaryotes, bacteria are less proficient in homologous recombination (HR) and non-homologous end joining (NHEJ). All existing genomic editing methods for Escherichia coli rely on exogenous HR or NHEJ systems to repair DNA double-strand breaks (DSBs). Although an E. coli native end-joining (ENEJ) system has been reported, its potential in chromosomal engineering has not yet been explored. Here, we present a CRISPR-Cas9-assisted native end-joining editing and show that ENEJ-dependent DNA repair can be used to conduct rapid and efficient knocking-out of E. coli genomic sequence of up to 83 kb. Moreover, the positive rate and editing efficiency is independent of high-efficiency competent cells. The method requires neither exogenous DNA repair systems nor introduced editing template. The Cas9 complex is the only foreign element in this method. This study is the first successful engineering effort to utilize ENEJ mechanism in genomic editing and provides an effective strategy for genetic engineering in bacteria that are inefficient in HR and NHEJ.
We report a novel method to freely transform the modes of a perfect optical vortex (POV). By adjusting the scaling factor of the Bessel-Gauss beam at the object plane, the POV mode transformation can be easily controlled from circle to ellipse with a high mode purity. Combined with the modulation of the cone angle of an axicon, the ellipse mode can be freely adjusted along the two orthogonal directions. The properties of the "perfect vortex" are experimentally verified. Moreover, fractional elliptic POVs with versatile modes are presented, where the number and position of the gaps are controllable. These findings are significant for applications that require the complex structured optical field of the POV.
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