Cloning, expression, crystallization and preliminary X‐ray studies of a superfolder GFP fusion of cyanobacterial Psb32. Issue 4 (1st April 2015)

• Record Type: Journal Article
• Authors: Liauw, Pasqual; Kannchen, Daniela; Gasper, Raphael; Dyczmons‐Nowaczyk, Nina; Nowaczyk, Marc M.; Hofmann, Eckhard
• Is Part Of: Acta crystallographica. Volume 71:Issue 4(2015:Apr.)
• Page Start: 409
• Page End: 413
• Publication Date: 2015-04-01
• Subjects: Crystallography -- Periodicals; Crystals -- Periodicals; 548
• DOI: 10.1107/S2053230X15003970
• Languages: English
• ISSNs: 2053-230X

Abstract

<abstract abstract-type="main" xml:lang="en"> <title> <x xml:space="preserve">Abstract</x> </title> <p>A fusion of Psb32 from the thermophilic cyanobacterium <italic>Thermosynechococcus elongatus</italic> BP‐1 (<italic>Te</italic>Psb32) with superfolder GFP was created for enhanced solubility and improved detection and purification. The fusion protein readily formed large hexagonal crystals belonging to space group <italic>P</italic>6₁22. A full data set extending to 2.3 Å resolution was collected at the Swiss Light Source. The phase problem could be solved by using only the sfGFP fusion partner or by using GFP and <italic>At</italic>TLP18.3 from <italic>Arabidopsis thaliana</italic> as search models. Based on this expression construct, a versatile library of 24 vectors combining four different superfolder GFP variants and three affinity tags was generated to facilitate expression and screening of fluorescent fusion proteins.</p> </abstract>