Kinetic characterization (Km and Vmax) of recombinant EiGS1-1 proteins was performed for the two substrates, glutamate and ATP, by the biosynthetic assay (Jalaludin et al., 2017) . The reaction mixture consisted of 100 mM Tris-HCl (pH 7.5), 10 mM ATP, 20 mM MgCl2, 30 mM hydroxylamine and 20 mM glutamate.
Glutamine Synthetase Enzyme Assay
Kinetic characterization (Km and Vmax) of recombinant EiGS1-1 proteins was performed for the two substrates, glutamate and ATP, by the biosynthetic assay (Jalaludin et al., 2017) . The reaction mixture consisted of 100 mM Tris-HCl (pH 7.5), 10 mM ATP, 20 mM MgCl2, 30 mM hydroxylamine and 20 mM glutamate.
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization : University of Western Australia
Other organizations : Guangdong Academy of Agricultural Sciences, Institute of Plant Protection, Taras Shevchenko National University of Kyiv
Variable analysis
- Glufosinate concentration (0, 0.001, 0.01, 0.025, 0.05, 0.1, 0.5, and 1 mM)
- GS activity in E. indica tissue extracts and in yeast recombinant EiGS1-1 proteins (measured by the γ-transferase assay)
- Kinetic characteristics (Km and Vmax) of recombinant EiGS1-1 proteins for the substrates glutamate and ATP (measured by the biosynthetic assay)
- Protein concentration (for sample calibration)
- Reaction mixture composition (100 mM Tris-HCl (pH 7.5), 10 mM ATP, 20 mM MgCl2, 30 mM hydroxylamine, and 20 mM glutamate)
- Positive control: Not specified
- Negative control: Not specified
Annotations
Based on most similar protocols
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!