Year: 2017
Type: article
Abstract: Originally published in PROTEINS: Structure, Function, and Bioinformatics 2015;83(5): 799–804(DOI: 10.1002/prot.24769) There was an error in the legend for figure 2. The correct legend is shown below. Figure 2. The CRP/CRPG reporter system for detecting adenylate and guanylate cyclase activities. A, The BL21 [DE3] cya crp strains expressing wild-type CRP from pRKcrp (A) or CRPG from pRKcrpG (G) were grown with or without the addition of cAMP or cGMP. The strain expressing CRP developed blue color only in the presence of cAMP whereas the strain expressing CRPG developed blue color in the presence of either cAMP or cGMP. B, Scheme of cAMP- and cGMP-dependent lacZ expression. C, Heterologous nucleotide cyclases were introduced into the BL21 [DE3] cya crp mutants expressing CRP or CRPG. The CyaB1 (catalytic domain) and CyaA (full-length) produced blue color in both strains, while GcyA, GuaA and GuaX produced blue colonies only in the presence of CRPG. The medium contained 50 μM IPTG. D, Analysis of secreted cNMPs from the E. coli strains expressing nucleotide cyclases. The same strains as in panel C were used. Cyclic nucleotide levels were measured by ELISA using cAMP- and cGMP-specific antibodies. Average data and standard deviations from three independent experiments are shown.
DOI: 10.1002/prot.25308
Authors: Min‐Hyung Ryu, Hwan Youn, In‐Hye Kang, Mark Gomelsky
Institutions: University of Wyoming, California State University, Fresno
Language: English
Subfield: Genetics
Domain: Life Sciences
Open Access status: bronze
OpenAlex ID: W2130065169