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Purpose: To determine the mechanism of plasma membrane binding and function of BFSP1 in this cell compartment.

Methods: Lens membranes were prepared by sequential extraction, culminating with a treatment with 0.1 M sodium hydroxide, a standard biochemical approach for the purification ofintegral membrane proteins. Full length human BFSP1 was obtained from Source Bioscience (http://www. lifesciences. sourcebioscience. com/IMAGE clones 6154051 and 5406467). The C-terminal region of HsBFSP1 was amplified by PCR, the products cloned into and sequenced in pGEM-T Easy (Promega, UK; www. promega. com). These were inserted into the following vector systems: pET23/28 retaining the C-terminal His tag for subsequent purification; peGFPN3 for expression in tissue culture cells by transient transfection; pXβG for expression in Xenopus oocytes for oocyte swelling assays. BFSP1 and its …