Bacteria cell sorting

J. Porter, C. Edwards, J. A. W. Morgan, and R. W. Pickup, Rapid, automated separation of specific bacteria from lake water and sewage by flow cytometry and cell sorting, Appl. Environ. Microbiol. 59 3327 (1993). 

Yeast display has been proposed recently as an alternative way to display mammalian proteins. Because some eukaryotic proteins expressed in E. coli are not in the soluble form, they cannot be incorporated into phage particles. Furthermore, phage display may sometimes select for reduced host toxicity or higher infectivity rather than increased affinity. Yeast display should alleviate expression biases present in E. coli and, as bacteria, have the advantage of high expression levels of displayed fusion polypeptides and selection through flow cytometric cell sorting. 

Porter,)., Robinson,)., Pickup, R. and Edwards, C.). (1998) An evaluation of lectin-mediated magnetic bead cell sorting for the targeted separation of enteric bacteria. Journal of Applied Microbiology, 84, ITl- il. 

The yields of DNA intake by the cells are, however, very low. The few bacteria containing the hybrid plasmid need to be sorted out. One technique applies a plasmid which encodes for antibiotic-degrading enzymes, e.g for penicillinase and tetracyclinase, and thus... 

As described in Section II,A, gram-positive bacteria have only one membrane (the cytoplasmic membrane). Therefore, the translocation through the Sec pathway directly leads proteins to be secreted (Simonen and Palva, 1993 Nagarajan, 1993). The issue of protein sorting into the cell wall is described in a separate section. 

In spite of the variety of appearances of eukaryotic cells, their intracellular structures are essentially the same. Because of their extensive internal membrane structure, however, the problem of precise protein sorting for eukaryotic cells becomes much more difficult than that for bacteria. Figure 4 schematically illustrates this situation. There are various membrane-bound compartments within the cell. Such compartments are called organelles. Besides the plasma membrane, a typical animal cell has the nucleus, the mitochondrion (which has two membranes see Fig. 6), the peroxisome, the ER, the Golgi apparatus, the lysosome, and the endosome, among others. As for the Golgi apparatus, there are more precise distinctions between the cis, medial, and trans cisternae, and the TGN trans Golgi network) (see Fig. 8). In typical plant cells, the chloroplast (which has three membranes see Fig. 7) and the cell wall are added, and the lysosome is replaced with the vacuole. 

Schneewind, O., Mihaylova-Petkov, D., and Model, P. (1993). Cell wall sorting signals in surface proteins of gram-positive bacteria. EMBO J. 12, 4803-4811. 

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