Protocol detail

Subcellular Localization of OsNPF7.6

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The coding sequence of OsNPF7.6 for subcellular localization was amplified using the pMD19-T vector and ligated into pSAT6A-GFP [33 (link)] with the correct direction. The OsNPF7.6-GFP vector was transformed into Arabidopsis mesophyll protoplasts and identified by a laser scanning microscope (LSM410; Carl Zeiss, Jena, Germany). Plasma membrane dye FMTM4-64FX (Invitrogen, Life Technologies, Carlsbad, CA, USA) was used as marker.

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Zhang M., Lai L., Liu X., Liu J., Liu R., Wang Y., Liu J, & Chen J. (2022). Overexpression of Nitrate Transporter 1/Peptide Gene OsNPF7.6 Increases Rice Yield and Nitrogen Use Efficiency. Life, 12(12), 1981.

Publication 2022

LSM410 FMTM4-64FX

Arabidopsis Coding sequence Laser scanning microscope Plasma membrane Protoplasts Vector

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Variable analysis

independent variables

Insertion of the coding sequence of OsNPF7.6 into the pSAT6A-GFP vector

dependent variables

Subcellular localization of OsNPF7.6-GFP fusion protein in Arabidopsis mesophyll protoplasts

control variables

Use of the pMD19-T vector to amplify the coding sequence of OsNPF7.6
Ligation of the OsNPF7.6 coding sequence into the pSAT6A-GFP vector in the correct direction
Transformation of the OsNPF7.6-GFP vector into Arabidopsis mesophyll protoplasts
Identification of the OsNPF7.6-GFP fusion protein using a laser scanning microscope (LSM410; Carl Zeiss, Jena, Germany)
Use of the plasma membrane dye FM4-64FX (Invitrogen, Life Technologies, Carlsbad, CA, USA) as a marker

controls

No positive or negative controls were explicitly mentioned in the provided information.

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