Biotechnology Bulletin ›› 2021, Vol. 37 ›› Issue (10): 196-202.doi: 10.13560/j.cnki.biotech.bull.1985.2021-0069
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ZHENG Fang-fang1(), LIN Jun-sheng2()
Received:
2021-01-17
Online:
2021-10-26
Published:
2021-11-12
Contact:
LIN Jun-sheng
E-mail:492969673@qq.com;junshenglin@hqu.edu.cn
ZHENG Fang-fang, LIN Jun-sheng. Selection and Specificity of Nucleic Acid Aptamers for a Proliferation Inducing Ligand[J]. Biotechnology Bulletin, 2021, 37(10): 196-202.
SELEX round | APRIL/μg | APRIL incubation time/min | BAFF/μg | BAFF incubation time/min | ssDNA library/pmol |
---|---|---|---|---|---|
1 | 6 | 60 | - | - | 1 000 |
2 | 4 | 45 | - | - | 200 |
3 | 4 | 45 | - | - | 200 |
4 | 4 | 45 | 2 | 30 | 200 |
5 | 2 | 30 | 2 | 30 | 200 |
6 | 2 | 30 | 2 | 30 | 200 |
7 | 2 | 30 | 2 | 30 | 200 |
Table 1 Optimization of the selection conditions of the APRIL aptamers
SELEX round | APRIL/μg | APRIL incubation time/min | BAFF/μg | BAFF incubation time/min | ssDNA library/pmol |
---|---|---|---|---|---|
1 | 6 | 60 | - | - | 1 000 |
2 | 4 | 45 | - | - | 200 |
3 | 4 | 45 | - | - | 200 |
4 | 4 | 45 | 2 | 30 | 200 |
5 | 2 | 30 | 2 | 30 | 200 |
6 | 2 | 30 | 2 | 30 | 200 |
7 | 2 | 30 | 2 | 30 | 200 |
Item | Sequence of random region(5'-3') | Round 4/% | Round 5/% | Round 6/% | Round 7/% | dG/ (kcal·mol-1) | Stem-loop structure | G/% |
---|---|---|---|---|---|---|---|---|
Apt 1 | GGTTGGGTTGGGGGGGTACTCTGAACTCGCATAGCCGGTG | 0.7 | 1.4 | 3.8 | 4.7 | -8.9 | 4 | 45.0 |
Apt 2 | AAGAGGCGGGCGGGTGGGGCTGACCTCGCACTTGGTTCTC | 0.3 | 1.5 | 2.5 | 5.2 | -8.2 | 5 | 42.5 |
Apt 3 | TGGTGGGAGGTATACGGGTGGTGGGTGGGGGTGTTGATGG | 0.1 | 3.1 | 3.3 | 3.4 | -7.5 | 5 | 60.0 |
Apt 5 | GATCCTCGCGTAGGCTCGGGGTGAGGTGGGTGGGTTCGTG | 0.6 | 2.2 | 2.9 | 3.7 | -10.0 | 4 | 50.0 |
Apt 7 | GGTCGGGGTGGGTGGGCGACCTCGCATAAGTTAGGCAGTT | 0.2 | 2.7 | 3.0 | 3.9 | -8.7 | 4 | 45.0 |
Apt 10 | ACTGGGGGGGGTCGGGTGGGTGTCTCGGACCTCGCATCA | 0.5 | 2.3 | 3.1 | 3.8 | -8.8 | 5 | 46.2 |
Apt 14 | TGTTGGGTTGGGGGGGTACTCTGAACTCGCATAGCCGGTG | 0.1 | 1.4 | 4.2 | 4.2 | -9.0 | 3 | 42.5 |
Apt 16 | GGTCGGGTGGGTGGGCGACCTCGCATAGCGATTGATGATC | 0.1 | 1.0 | 4.2 | 4.6 | -11.5 | 3 | 42.5 |
Table 2 Sequences of candidate aptamers
Item | Sequence of random region(5'-3') | Round 4/% | Round 5/% | Round 6/% | Round 7/% | dG/ (kcal·mol-1) | Stem-loop structure | G/% |
---|---|---|---|---|---|---|---|---|
Apt 1 | GGTTGGGTTGGGGGGGTACTCTGAACTCGCATAGCCGGTG | 0.7 | 1.4 | 3.8 | 4.7 | -8.9 | 4 | 45.0 |
Apt 2 | AAGAGGCGGGCGGGTGGGGCTGACCTCGCACTTGGTTCTC | 0.3 | 1.5 | 2.5 | 5.2 | -8.2 | 5 | 42.5 |
Apt 3 | TGGTGGGAGGTATACGGGTGGTGGGTGGGGGTGTTGATGG | 0.1 | 3.1 | 3.3 | 3.4 | -7.5 | 5 | 60.0 |
Apt 5 | GATCCTCGCGTAGGCTCGGGGTGAGGTGGGTGGGTTCGTG | 0.6 | 2.2 | 2.9 | 3.7 | -10.0 | 4 | 50.0 |
Apt 7 | GGTCGGGGTGGGTGGGCGACCTCGCATAAGTTAGGCAGTT | 0.2 | 2.7 | 3.0 | 3.9 | -8.7 | 4 | 45.0 |
Apt 10 | ACTGGGGGGGGTCGGGTGGGTGTCTCGGACCTCGCATCA | 0.5 | 2.3 | 3.1 | 3.8 | -8.8 | 5 | 46.2 |
Apt 14 | TGTTGGGTTGGGGGGGTACTCTGAACTCGCATAGCCGGTG | 0.1 | 1.4 | 4.2 | 4.2 | -9.0 | 3 | 42.5 |
Apt 16 | GGTCGGGTGGGTGGGCGACCTCGCATAGCGATTGATGATC | 0.1 | 1.0 | 4.2 | 4.6 | -11.5 | 3 | 42.5 |
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