Biotechnology Bulletin ›› 2024, Vol. 40 ›› Issue (5): 66-73.doi: 10.13560/j.cnki.biotech.bull.1985.2023-1178
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ZHANG Zhen1(), LI Qing2, XU Jing2, CHEN Kai-yuan2, ZHANG Chun-zhi2, ZHU Guang-tao1()
Received:
2023-12-13
Online:
2024-05-26
Published:
2024-06-13
Contact:
ZHU Guang-tao
E-mail:zzhen1181@163.com;zhuguangtao@ynnu.edu.cn
ZHANG Zhen, LI Qing, XU Jing, CHEN Kai-yuan, ZHANG Chun-zhi, ZHU Guang-tao. Construction and Application of Potato Mitochondrial Targeted Expression Vector[J]. Biotechnology Bulletin, 2024, 40(5): 66-73.
扩增片段Amplified fragment | 引物序列Primer sequence(5'-3') | ||
---|---|---|---|
2×35S启动子组合 | 2×35S | F | TATGACCATGATTACGAATTCGCTAGAGCAGCTTGCCAACATG |
2×35S | R | CCATTGCCATGGATCCAAGCTTGAAGAGAGAGAC | |
2×35S | R* | GGCGTGCCATGGTCGATCGACAGATCTGCG | |
ATPγ | F | TCGATCGACCATGGCAATGGCTGTTTTCCG | |
ATPγ | R | TGCTCACCATGCCTTGAACTGCTCTAAGCTTGG | |
Rf1b | F | TCGATCGACCATGGCACGCCGCGTCGCTG | |
Rf1b | R | TGCTCACCATGCGGTTGAAGCGGGACAC | |
eGFP | F | AGTTCAAGGCATGGTGAGCAAGGGCGAGG | |
eGFP | F* | CTTCAACCGCATGGTGAGCAAGGGCGAGG | |
eGFP | R | ACGACGGCCAGTGCCAAGCTTGATCTAGTAACATAGATGACACCGC | |
StUBI10启动子组合 | StUBI10 | F | TATGACCATGATTACGAATTCCCAAGACAATTTCAGCTTAAAAAGT |
StUBI10 | R | TGCTCACCATGCCTTGAACTGCTCTAAGCTTGG | |
StUBI10 | R* | CGCGGCGTGCCATGGATCCAAGCTTGAATCTGCAAATTC | |
ATPγ | F | GCTTGGATCCATGGCAATGGCTGTTTTCCG | |
ATPγ | R | TGCTCACCATGCCTTGAACTGCTCTAAGCTTGG | |
Rf1b | F | GCTTGGATCCATGGCACGCCGCGTCGCTG | |
Rf1b | R | TGCTCACCATGCGGTTGAAGCGGGACAC | |
eGFP | F | AGTTCAAGGCATGGTGAGCAAGGGCGAGG | |
eGFP | F* | CTTCAACCGCATGGTGAGCAAGGGCGAGG | |
eGFP | R | ACGACGGCCAGTGCCAAGCTTGATCTAGTAACATAGATGACACCGC | |
AtUBI10启动子组合 | AtUBI10 | F | TATGACCATGATTACGAATTCATACGCTTCAATGCAGTGG |
AtUBI10 | R | CCATTGCCATTGATAACTCTAGAATCTGTTAATCA | |
AtUBI10 | R* | GGCGTGCCATTGATAACTCTAGAATCTGTTAATCA | |
ATPγ | F | AGAGTTATCAATGGCAATGGCTGTTTTCCG | |
ATPγ | R | TGCTCACCATGCCTTGAACTGCTCTAAGCTTGG | |
Rf1b | F | AGAGTTATCAATGGCACGCCGCGTCGCTG | |
Rf1b | R | TGCTCACCATGCGGTTGAAGCGGGACAC | |
eGFP | F | AGTTCAAGGCATGGTGAGCAAGGGCGAGG | |
eGFP | F* | CTTCAACCGCATGGTGAGCAAGGGCGAGG | |
eGFP | R | ACGACGGCCAGTGCCAAGCTTGATCTAGTAACATAGATGACACCGC | |
P2300 | F | GTTAGCTCACTCATTAGGCAC | |
P2300 | R | GCTGGCGTAATAGCGAAGAG |
Table 1 PCR primer sequences used in this study
扩增片段Amplified fragment | 引物序列Primer sequence(5'-3') | ||
---|---|---|---|
2×35S启动子组合 | 2×35S | F | TATGACCATGATTACGAATTCGCTAGAGCAGCTTGCCAACATG |
2×35S | R | CCATTGCCATGGATCCAAGCTTGAAGAGAGAGAC | |
2×35S | R* | GGCGTGCCATGGTCGATCGACAGATCTGCG | |
ATPγ | F | TCGATCGACCATGGCAATGGCTGTTTTCCG | |
ATPγ | R | TGCTCACCATGCCTTGAACTGCTCTAAGCTTGG | |
Rf1b | F | TCGATCGACCATGGCACGCCGCGTCGCTG | |
Rf1b | R | TGCTCACCATGCGGTTGAAGCGGGACAC | |
eGFP | F | AGTTCAAGGCATGGTGAGCAAGGGCGAGG | |
eGFP | F* | CTTCAACCGCATGGTGAGCAAGGGCGAGG | |
eGFP | R | ACGACGGCCAGTGCCAAGCTTGATCTAGTAACATAGATGACACCGC | |
StUBI10启动子组合 | StUBI10 | F | TATGACCATGATTACGAATTCCCAAGACAATTTCAGCTTAAAAAGT |
StUBI10 | R | TGCTCACCATGCCTTGAACTGCTCTAAGCTTGG | |
StUBI10 | R* | CGCGGCGTGCCATGGATCCAAGCTTGAATCTGCAAATTC | |
ATPγ | F | GCTTGGATCCATGGCAATGGCTGTTTTCCG | |
ATPγ | R | TGCTCACCATGCCTTGAACTGCTCTAAGCTTGG | |
Rf1b | F | GCTTGGATCCATGGCACGCCGCGTCGCTG | |
Rf1b | R | TGCTCACCATGCGGTTGAAGCGGGACAC | |
eGFP | F | AGTTCAAGGCATGGTGAGCAAGGGCGAGG | |
eGFP | F* | CTTCAACCGCATGGTGAGCAAGGGCGAGG | |
eGFP | R | ACGACGGCCAGTGCCAAGCTTGATCTAGTAACATAGATGACACCGC | |
AtUBI10启动子组合 | AtUBI10 | F | TATGACCATGATTACGAATTCATACGCTTCAATGCAGTGG |
AtUBI10 | R | CCATTGCCATTGATAACTCTAGAATCTGTTAATCA | |
AtUBI10 | R* | GGCGTGCCATTGATAACTCTAGAATCTGTTAATCA | |
ATPγ | F | AGAGTTATCAATGGCAATGGCTGTTTTCCG | |
ATPγ | R | TGCTCACCATGCCTTGAACTGCTCTAAGCTTGG | |
Rf1b | F | AGAGTTATCAATGGCACGCCGCGTCGCTG | |
Rf1b | R | TGCTCACCATGCGGTTGAAGCGGGACAC | |
eGFP | F | AGTTCAAGGCATGGTGAGCAAGGGCGAGG | |
eGFP | F* | CTTCAACCGCATGGTGAGCAAGGGCGAGG | |
eGFP | R | ACGACGGCCAGTGCCAAGCTTGATCTAGTAACATAGATGACACCGC | |
P2300 | F | GTTAGCTCACTCATTAGGCAC | |
P2300 | R | GCTGGCGTAATAGCGAAGAG |
Fig. 2 Colocalization of eGFP and mitochondria in transient tobacco infection A1 is the vector AtUBI10::ATPγ-eGFP, A2 is the vector AtUBI10::Rf1b-eGFP; B1 is the vector StUBI10::ATPγ-eGFP, B2 is the vector StUBI10::Rf1b-eGFP; C1 is the carrier 2×35S::ATPγ-eGFP, C2 is the carrier 2×35S::Rf1b-eGFP. Mitochondrial localization marker by mCherry. The same below
Fig. 3 Fluorescence intensity statistics of tobacco transient infestation and potato protoplast transformation A is the average fluorescence value statistics of eGFP and mCherry for each vector in tobacco transient infestation, and B is the average fluorescence value statistics of eGFP and mCherry for each vector in potato protoplast transformation. Data are in mean±SD(n=3). ns indicates no significant differences according to student's t-test, and *(P<0.05)indicates significant differences according to student's t-test
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