Preparation of Electro-competent Bacterial Cells for Transformation
2:33
Electroporation of Bacterial Cells with Complementation Plasmids
3:39
Transformation of AOH1 to Facilitate Functional Complementation Using L,L-diaminopimelate Aminotransferase
4:18
Transformation of TKL-11 to Facilitate Functional Complementation Using UDP-N-acetylmuramoylalanyl-D-glutamyl-2,6-meso-diaminopimelate Ligase
5:12
Transformation of Mutant Hx699 to Facilitate Functional Complementation of Hypomucoid Phenotype of Novosphingobium sp. Strain Hx699
6:02
Results: Functional Complementation Analysis
7:44
Conclusion
Transcrição
The overall goal of the Functional Complementation Essay is to elucidate the activity of an enzyme by introducing a functional copy of the corresponding gene into a mutant cell to see if it restores a wild type phenotype in the mutant background.
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The validation of enzymatic activities involved in biochemical pathways can be elucidated using functional complementation analysis (FCA). Described in this manuscript is the FCA assay demonstrating the enzymatic activity of enzymes involved in the metabolism of amino acids, bacterial stringent response and bacterial peptidoglycan biosynthesis.