Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (10): 254-261.doi: 10.13560/j.cnki.biotech.bull.1985.2022-0015
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ZHANG Hao1,2(), HE Chang-sheng1,2, LI Yan-yan1,2, WANG Yong1,2(), ZHU Jiang-jiang2, Emu Quzhe3, LIN Ya-qiu1,2()
Received:
2022-01-05
Online:
2022-10-26
Published:
2022-11-11
Contact:
WANG Yong,LIN Ya-qiu
E-mail:zhanghao9235@163.com;wangyong010101@swun.cn;linyq1999@163.com
ZHANG Hao, HE Chang-sheng, LI Yan-yan, WANG Yong, ZHU Jiang-jiang, Emu Quzhe, LIN Ya-qiu. Regulation of miR-301b on Goat Intramuscular Adipocyte Differentiation[J]. Biotechnology Bulletin, 2022, 38(10): 254-261.
名称Name | 序列Sequence(5'-3') |
---|---|
miR-301b mimics | CAGUGCAAUGAUAUUGUCAAAGC UUUGACAAUAUCAUUGCACUGUU |
mimics NC | UUCUCCGAACGUGUCACGUTTACGUGACACG- UUCGGAGAATT |
miR-301b Inhibitor | GCUUUGACAAUAUCAUUGCACUG |
Inhibitor NC | CAGUACUUUUGUGUAGUACAA |
Table 1 Sequences of miRNA mimic and inhibitor
名称Name | 序列Sequence(5'-3') |
---|---|
miR-301b mimics | CAGUGCAAUGAUAUUGUCAAAGC UUUGACAAUAUCAUUGCACUGUU |
mimics NC | UUCUCCGAACGUGUCACGUTTACGUGACACG- UUCGGAGAATT |
miR-301b Inhibitor | GCUUUGACAAUAUCAUUGCACUG |
Inhibitor NC | CAGUACUUUUGUGUAGUACAA |
基因 Gene/miRNA | 序列 Sequence(5'-3') | Tm/℃ | 产物长度 Product length/ bp | 登录号 GenBank No. |
---|---|---|---|---|
AP2 | TGAAGTCACTCCAGATGACAGGTGACACATTCCAGCACCAGC | 58 | 143 | NM_001285623.1 |
CEBPα | CCGTGGACAAGAACAGCAACAGGCGGTCATTGTCACTGGT | 58 | 142 | XM_018062278.1 |
CEBPβ | CAAGAAGACGGTGGACAAGCAACAAGTTCCGCAGGGTG | 66 | 204 | XM_018058020.1 |
PPARγ | AAGCGTCAGGGTTCCACTATG GAACCTGATGGCGTTATGAGAC | 60 | 197 | NM_001285658.1 |
Pref1 | CCGGCTTCATGGATAAGACCTGCCTCGCACTTGTTGAGGAA | 65 | 184 | KP686197.1 |
SREBP1 | AAGTGGTGGGCCTCTCTGA GCAGGGGTTTCTCGGACT | 58 | 127 | NM_001285755.1 |
KLF3 | CTCGGTGTCATACCCGTCTAATCCATCTGCATCCCGTGAGAC | 60 | 128 | KU041753.1 |
UXT | GCAAGTGGATTTGGGCTGTAACATGGAGTCCTTGGTGAGGTTGT | 60 | 180 | XM_005700842.2 |
U6 | TGGAACGCTTCACGAATTTGCGGGAACGATACAGAGAAGATTAGC | 60 | ||
miR-301b | CTGGAGCAGTGCAATGATGAAAGTGCAGGGTCCGAGGT | 58 |
Table 2 Sequences information of specific primers
基因 Gene/miRNA | 序列 Sequence(5'-3') | Tm/℃ | 产物长度 Product length/ bp | 登录号 GenBank No. |
---|---|---|---|---|
AP2 | TGAAGTCACTCCAGATGACAGGTGACACATTCCAGCACCAGC | 58 | 143 | NM_001285623.1 |
CEBPα | CCGTGGACAAGAACAGCAACAGGCGGTCATTGTCACTGGT | 58 | 142 | XM_018062278.1 |
CEBPβ | CAAGAAGACGGTGGACAAGCAACAAGTTCCGCAGGGTG | 66 | 204 | XM_018058020.1 |
PPARγ | AAGCGTCAGGGTTCCACTATG GAACCTGATGGCGTTATGAGAC | 60 | 197 | NM_001285658.1 |
Pref1 | CCGGCTTCATGGATAAGACCTGCCTCGCACTTGTTGAGGAA | 65 | 184 | KP686197.1 |
SREBP1 | AAGTGGTGGGCCTCTCTGA GCAGGGGTTTCTCGGACT | 58 | 127 | NM_001285755.1 |
KLF3 | CTCGGTGTCATACCCGTCTAATCCATCTGCATCCCGTGAGAC | 60 | 128 | KU041753.1 |
UXT | GCAAGTGGATTTGGGCTGTAACATGGAGTCCTTGGTGAGGTTGT | 60 | 180 | XM_005700842.2 |
U6 | TGGAACGCTTCACGAATTTGCGGGAACGATACAGAGAAGATTAGC | 60 | ||
miR-301b | CTGGAGCAGTGCAATGATGAAAGTGCAGGGTCCGAGGT | 58 |
Fig.1 Effect of overexpressing miR-301b on the morphol-ogy of goat intramuscular adipocytes A:The overexpressing efficiency of miR-301b. B:Oil red O staining and Bodipy staining(×400). C:OD value of oil red O staining. D:Triglyceride(TG)content assay. * indicates P<0.05, ** indicates P<0.01, the same below
Fig. 3 Effect of interfering miR-301b on the morphology of goat intramuscular adipocytes A:Efficiency of miR-301b. B:Oil red O staining and Bodipy staining(×400).C:OD value of oil red O staining. D:Triglyceride(TG)content assay
Fig. 5 Prediction and expression level detection of miR-301b target gene A:miR-301b target gene prediction Venn diagram;B:KLF3 potential miRNA binding site prediction;C:KLF3 expression level after simulating miR-301b expression;D:KLF3 expression level after interfering with miR-301b expression
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