Biotechnology Bulletin ›› 2023, Vol. 39 ›› Issue (9): 168-175.doi: 10.13560/j.cnki.biotech.bull.1985.2023-0150
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LIU Yu-ling1,2(), WANG Meng-yao2, SUN Qi1, MA Li-hua1, ZHU Xin-xia1()
Received:
2023-02-22
Online:
2023-09-26
Published:
2023-10-24
Contact:
ZHU Xin-xia
E-mail:839779394@qq.com;xinxiazh@shzu.edu.cn
LIU Yu-ling, WANG Meng-yao, SUN Qi, MA Li-hua, ZHU Xin-xia. Effect of RD29A Promoter on the Stress Resistance of Transgenic Tobacco with SikCDPK1 Gene from Saussurea involucrata[J]. Biotechnology Bulletin, 2023, 39(9): 168-175.
引物Primer name | 引物序列Primer sequence(5'-3') | 用途Purpose |
---|---|---|
RD29A-F | GAATTCCGACTCAAAACAAACTTACGAA | 启动子克隆Promoter cloing |
RD29A-R | CCCGGGAATCAAACCCTTTATTCCTGA | 启动子克隆Promoter cloing |
SikCDPK1-F | GGATCCATGGGGAATACTTGTGTTGGAC | 基因鉴定 Gene identification |
SikCDPK1-R | GTCGACCCGTCGATACCGGAAAAAAC | 基因鉴定 Gene identification |
Table 1 Primers information
引物Primer name | 引物序列Primer sequence(5'-3') | 用途Purpose |
---|---|---|
RD29A-F | GAATTCCGACTCAAAACAAACTTACGAA | 启动子克隆Promoter cloing |
RD29A-R | CCCGGGAATCAAACCCTTTATTCCTGA | 启动子克隆Promoter cloing |
SikCDPK1-F | GGATCCATGGGGAATACTTGTGTTGGAC | 基因鉴定 Gene identification |
SikCDPK1-R | GTCGACCCGTCGATACCGGAAAAAAC | 基因鉴定 Gene identification |
Fig. 1 Double enzyme digestion of RD29A∷SikCDPK1 recombinant plasmid A: Recombinant plasmids was digested with double enzymes(M: Marker III.1: Plasmid control. 2: Digestion products). B: Recombinant plasmids were digested with double enzymes(M: Trans 5 K DNA marker. 1-2: Digestion products.3: Plasmid control)
Fig. 2 Identification of transgenic tobacco by PCR A: PCR identification of 35S∷SikCDPK1 transgenic tobacco(M: DL2000 DNA marker. 1-17: Transgenic plant). B: PCR detection of RD29A∷SikCDPK1 transgenic tobacco SikCDPK1 gene(M: DL2000 DNA marker. 1-7: Detection of SikCDPK1 gene PCR. 8-16: WT PCR control). C: PCR detection of RD29A∷SikCDPK1 transgenic tobacco RD29A promoter(M: Marker III. 1-7: RD29A promoter PCR detection. 8: WT PCR control)
Fig. 3 Phenotypic characteristics of WT tobacco and transgenic tobacco under stress A, C: Normally growing tobacco. B: Withheld watering 30 d. D: Low temperature treatment of tobacco after 48 h. Line 1, Line 2: Two different tobacco lines
Fig. 4 Physiological indexes of non-transgenic tobacco and transgenic tobacco determined before and after stress * and * * showed significant correlations at 0.05 and 0.01 levels, respectively
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