The Application of <i>Nicotiana benthamiana</i> as a Transient Expression Host to Clone the Coding Sequences of Plant Genes
- 1 Key Laboratory of Chronic Diseases, Fuzhou Medical University, Fuzhou, China
- 2 Key Laboratory of Chronic Diseases, Fuzhou Medical University, Fuzhou, China
- 3 Key Laboratory of Chronic Diseases, Fuzhou Medical University, Fuzhou, China
- 4 Key Laboratory of Chronic Diseases, Fuzhou Medical University, Fuzhou, China
- 5 Key Laboratory of Chronic Diseases, Fuzhou Medical University, Fuzhou, China
- 6 Key Laboratory of Chronic Diseases, Fuzhou Medical University, Fuzhou, China
Abstract
Coding sequences (CDS) are commonly used for transient gene expression, in yeast two-hybrid screening, to verify protein interactions and in prokaryotic gene expression studies. CDS are most commonly obtained using complementary DNA (cDNA) derived from messenger RNA (mRNA) extracted from plant tissues and generated by reverse transcription. However, some CDS are difficult to acquire through this process as they are expressed at extremely low levels or have specific spatial and/or temporal expression patterns <i>in vivo</i>. These challenges require the development of alternative CDS cloning technologies. In this study, we found that the genomic intron-containing gene coding sequences (gDNA) from <i>Arabidopsis thaliana</i>, <i>Oryza sativa</i>, <i>Brassica napus</i>, and <i>Glycine max</i> can be correctly transcribed and spliced into mRNA in <i>Nicotiana benthamiana</i>. In contrast, gDNAs from <i>Triticum aestivum</i> and <i>Sorghum bicolor</i> did not function correctly. In transient expression experiments, the target DNA sequence is driven by a constitutive promoter. Theoretically, a sufficient amount of mRNA can be extracted from the <i>N. benthamiana</i> leaves, making it conducive to the cloning of CDS target genes. Our data demonstrate that <i>N. benthamiana</i> can be used as an effective host for the cloning CDS of plant genes.
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