@Article{biocell.2022.020229,
AUTHOR = {FENG ZHU, QINQIN ZHANG, YANGKAI ZHOU, QIPING ZHANG, MENGYAO CAO, ZHAOLIN JI},
TITLE = {Effects of two vectors on the expression of the NbNAC1 transcription factor and preparation of its polyclonal antibody},
JOURNAL = {BIOCELL},
VOLUME = {46},
YEAR = {2022},
NUMBER = {9},
PAGES = {2123--2131},
URL = {http://www.techscience.com/biocell/v46n9/47748},
ISSN = {1667-5746},
ABSTRACT = {The NAC (NAM, ATAF, and CUC) superfamily is one of the largest plant-specific families containing transcription factors. An increasing number of studies suggest that NAC1 is involved in plants response to different biotic and abiotic stimulis. <i>Nicotiana benthamiana</i> is a widely used system for evaluating plant-pathogen interactions. In order to study the biochemical function of NbNAC1, NbNAC1 protein and antibody are essential. Therefore, we focused on developing a prokaryotic expression system for producing the <i>Nicotiana benthamiana</i> NbNAC1 protein of in <i>Escherichia coli</i> and the preparation of its polyclonal antibody. Firstly, we constructed two different molecular weight prokaryotic expression vectors: pGE vector with GST tag (pGEX4T-1–NbNAC1) and pET expression vector with His tag (pET28a-NbNAC1). The NbNAC1 protein can be successfully expressed in both vectors. The His-tagged NbNAC1 proteins are insoluble, while the GST-tagged NbNAC1 proteins are partially soluble. We then successfully purified and enriched both proteins. The His-tagged NbNAC1 was chosen to immunize rabbits owing to an unknown protein accompanying the GST-tagged NbNAC1. The anti-NbNAC1 polyclonal antibody had good specificity and could be used in subsequent protein-related studies.},
DOI = {10.32604/biocell.2022.020229}
}