An Optimized Protocol for Detecting Guard Cell-specific Gene Expression by in situ RT-PCR in Brassica rapa .
| Autor: | Song Y; College of Horticulture, Henan Agricultural University, Zhengzhou, China.; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China., Guo X; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China., Wu J; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China., Liang J; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China., Lin R; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China., Yan Z; College of Horticulture, Henan Agricultural University, Zhengzhou, China., Wang X; Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, China. |
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| Jazyk: | angličtina |
| Zdroj: | Bio-protocol [Bio Protoc] 2023 Sep 05; Vol. 13 (17), pp. e4810. Date of Electronic Publication: 2023 Sep 05 (Print Publication: 2023). |
| DOI: | 10.21769/BioProtoc.4810 |
| Abstrakt: | Since the genetic transformation of Chinese cabbage ( Brassica rapa ) has not been well developed, in situ RT-PCR is a valuable option for detecting guard cell-specific genes. We reported an optimized protocol of in situ RT-PCR by using a FAMA homologous gene Bra001929 in Brassica rapa. FAMA in Arabidopsis has been verified to be especially expressed in guard cells. We designed specific RT-PCR primers and optimized the protocol in terms of the (a) reverse transcription time, (b) blocking time, (c) antigen-antibody incubation time, and (d) washing temperature. Our approach provides a sensitive and effective in situ RT-PCR method that can detect low-abundance transcripts in cells by elevating their levels by RT-PCR in the guard cells in Brassica rapa. Competing Interests: Competing interestsThe authors declare no conflicts of interest. (©Copyright : © 2023 The Authors; This is an open access article under the CC BY-NC license.) |
| Databáze: | MEDLINE |
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