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SpheroplastA spheroplast is a cell from which the cell wall has been almost completely removed, as by the action of penicillin. The name stems from the fact that after a microbe's cell wall is digested, membrane tension causes the cell to acquire a characteristic spherical shape. Spheroplasts are osmotically fragile, and will lyse if transferred to a hypotonic solution. Additional recommended knowledge
Uses and applicationsPatch clampingSpecially prepared giant spheroplasts of Gram-negative bacteria can be used to study the function of bacterial ion channels through a technique called patch clamp, which was originally designed for characterizing the behavior of neurons and other excitable cells. To prepare giant spheroplasts, bacteria are grown in a medium containing chemicals that prevent the cells from dividing completely. This causes bacteria to form long "snakes" that share a single membrane and cytoplasm. After a period of time, the cell walls of the "snakes" are digested, and the bacteria collapse into very large spheres surrounded by a single lipid bilayer. The membrane can then be analyzed on a patch clamp apparatus to determine the phenotype of the ion channels embedded in it. It is also common to overexpress a particular channel to amplify its effect and make it easier to characterize. The technique of patch clamping giant E. coli spheroplast have been used extensively for studying the native mechanosensitive channels (MscL, MscS, and MscM) of E. coli since 1987.[1][2] Recently, it has been extended to study other heterologously expressed ion channels and shown that it can be used as an ion-channel expression system comparable to Xenopus oocyte.[3][4][5] Cell lysisYeast cells are normally protected by a thick cell wall which makes extraction of cellular proteins difficult. Enzymatic digestion of the cell wall with zymolyase, creating spheroplasts, renders the cells vulnerable to easy lysis with detergents or rapid osmolar pressure changes. References
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This article is licensed under the GNU Free Documentation License. It uses material from the Wikipedia article "Spheroplast". A list of authors is available in Wikipedia. |