Biotechnology Bulletin ›› 2022, Vol. 38 ›› Issue (5): 175-182.doi: 10.13560/j.cnki.biotech.bull.1985.2021-1039
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LI Yu1(), CHEN Gang1(), MA Qian1(), KUANG Jie-hua1, CHEN You-ming2
Received:
2021-08-16
Online:
2022-05-26
Published:
2022-06-10
Contact:
CHEN Gang,MA Qian
E-mail:1948463705@qq.com;cheng@gdou.edu.cn;maq@gdou.edu.cn
LI Yu, CHEN Gang, MA Qian, KUANG Jie-hua, CHEN You-ming. Cloning of nanos1 Gene of Rachycentron canadum and Its Expression Analysis in the Embryonic and Gonadal Development[J]. Biotechnology Bulletin, 2022, 38(5): 175-182.
引物名称 Primer | 序列Sequence(5'-3') | 用途 Usage |
---|---|---|
Rcnanos1-F | ATGGATTTTCTCAACCACAACTATT | CDS序列克隆 CDS sequence cloning |
Rcnanos1-R | TTAGAATATTTTCATCCTCTTACCACC | |
Rcnanos1-3' F1 | GCCCAAAATCTGCGTCTTCTGC | 3' 末端序列克隆 3' RACE |
Rcnanos1-3' F2 | TCAAAAGACCAGCCATCCCAGC | |
Rcnanos1-5' R1 | GCTCCTGGAAGCGGGTTTGC | 5' 末端序列克隆 5' RACE |
Rcnanos1-5' R2 | CGATTACGCACGGACACGCC | |
Rcnanos1-F1 | AGAACCCCAACTCCATCACC | Rcnanos1的qRT-PCR检测 Expression ofRcnanos1 |
Rcnanos1-R1 | TCTGGAAGGGGCTCAGTATC | |
β-actin-F | AGGGAAATTGTGCGTGAC | 内参基因 Internal control gene |
β-actin-R | AGGCAGCTCGTAGCTCTT |
Table 1 Primer sequences used in this study
引物名称 Primer | 序列Sequence(5'-3') | 用途 Usage |
---|---|---|
Rcnanos1-F | ATGGATTTTCTCAACCACAACTATT | CDS序列克隆 CDS sequence cloning |
Rcnanos1-R | TTAGAATATTTTCATCCTCTTACCACC | |
Rcnanos1-3' F1 | GCCCAAAATCTGCGTCTTCTGC | 3' 末端序列克隆 3' RACE |
Rcnanos1-3' F2 | TCAAAAGACCAGCCATCCCAGC | |
Rcnanos1-5' R1 | GCTCCTGGAAGCGGGTTTGC | 5' 末端序列克隆 5' RACE |
Rcnanos1-5' R2 | CGATTACGCACGGACACGCC | |
Rcnanos1-F1 | AGAACCCCAACTCCATCACC | Rcnanos1的qRT-PCR检测 Expression ofRcnanos1 |
Rcnanos1-R1 | TCTGGAAGGGGCTCAGTATC | |
β-actin-F | AGGGAAATTGTGCGTGAC | 内参基因 Internal control gene |
β-actin-R | AGGCAGCTCGTAGCTCTT |
Fig.1 Complete sequence of Rcnanos1 cDNA and analysis of deduced amino acid sequence The initiation codon ATG and stop codon TAA are in bold. The polyadenylation signal ATTAAA are in bold and italic. The zinc finger motif are highlighted in grey shadow
Fig.3 Phylogenetic tree of nanos1 amino acid sequences based on Neighbor-Joining(NJ)method Numbers indicates the confidence level of each branch,the scale bar 0.05 in terms of genetic distance is indicated below the tree
Fig.4 Expressions of Rcnanos1 in the tissues of R. canadum by qRT-PCR LI:Liver. SP:Spleen. KI:Kidney. BR:Brain. HE:Heart. GI:Gill. TE:Testis. OV:Ovary. ST:Stomach. IN:Intestines. MU:Muscle. SK:Skin. EY:Eye. Different superscripts indicate significant difference(P<0.05)
Fig.5 Expressions of Rcnanos1 during the embryonic deve-lopment stages 1:1-cell stage;2:2-cell stage;3:4-cell stage;4:8-cell stage;5:16-cell stage;6:poly-cell stage;7:blastocyst stage;8:gastrulation stage;9:neuroblast stage;10:organ formation stage;11:hatching stage;12:1 day after hatching. Different superscripts indicate significant difference(P<0.05)
Fig.6 Rcnanos1 mRNA expression in annual gonadal deve-lopment of R. Canadum A:Rcnanos1 mRNA expression in testis;B:Rcnanos1 mRNA expression in ovary. Different superscripts indicate significant difference(P<0.05)
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