Abstract: Fluorescent fusion proteins were expressed in Bacillus cereus to visualize the germino-some by introducing a plasmid that carries fluorescent fusion proteins of germinant receptor GerR subunits or germinosome scaffold protein GerD. The effects of plasmid insertion and re-combinant protein expression on the spore proteome were investigated. Proteomic analysis showed that overexpression of the target proteins had negligible effects on the spore proteome. However, plasmid-bearing spores displayed dramatic abundance changes of spore proteins in-volved in signaling and metabolism. Our findings indicate that introduction of a plasmid alone alters spore protein composition dramatically, with 993 proteins significantly down-regulated and 415 proteins significantly up-regulated among 3323 identified proteins. This shows that empty vector controls are more appropriate to compare proteome changes due to plasmid en-coded genes than is the wild-type strain, when using plasmid based genetic tools. Therefore, re-searchers should keep in mind that molecular cloning techniques can alter more than their in-tended targets in a biological system, and interpret results with this in mind.
[doi:10.25345/C5PG1HS5V]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: Proteomics ; Bacillus cereus ; Spores ; Molecular Biology ; Plasmid ; Overexpression ; Germinosome
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Gertjan Kramer, University of Amsterdam, Netherlands |
Submitting User: | gkramer |
Xiaowei Gao, Bhagyashree N. Swarge, Winfried Roseboom, Yan Wang, Henk L. Dekker, Peter Setlow, Stanley Brul and Gertjan Kramer.
settings Open AccessArticle Changes in the Spore Proteome of Bacillus cereus in Response to Introduction of Plasmids.
Microorganisms 2022, 10(9), 1695.
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