Identification and Characterization of the DMRT11E Gene in the Oriental River Prawn Macrobrachium nipponense
Abstract
:1. Introduction
2. Results
2.1. Molecular Cloning and Structural Analysis of DMRT11E Gene
2.2. Tissue-Specific Expression Patterns of MniDMRT11E
2.3. Expression of the MniDMRT11E Gene During Embryo Stages
2.4. Expression of the MniDMRT11E Gene in Different Developmental Stages of the Ovaries
2.5. Localization of the MniDMRT11E Gene in the Gonad
2.6. Effects of MniDMRT11E Knockdown on Gonad by RNAi
3. Discussion
4. Materials and Methods
4.1. Experimental Animals and Sampling
4.2. Nucleotide Sequence and Bioinformatics Analysis
4.3. Tissue Expression Analysis by Quantitative Real-Time PCR
4.4. Expression Profiles of DMRT11E in Ovarian Cycle
4.5. In Situ Hybridization (ISH)
4.6. RNA Interference of DMRT11E
4.7. Data Analysis
Author Contributions
Funding
Conflicts of Interest
Abbreviations
DMRT | doublesex and mab-3 related transcription factor |
VG | vitellogenin |
IAG | insulin-like androgenic gland factor |
dsx | doublesex |
qPCR | quantitative real-time reverse transcription PCR |
EM | eosinophilic matrix |
E | eyestalk |
BR | brain |
H | heart |
HE | hepatopancreas |
G | gill |
M | muscle; |
O | ovary |
T | testis |
AG | abdominal ganglia |
UE | unfertilized egg |
CS | cleavage stage |
BS | blastula stage |
GS | gastrula stage |
NS | nauplius stage |
ZS | zoea stage |
L1 | the first day larvae after hatching |
O I | undeveloped stage |
O II | developing stage |
O III | nearly-ripe stage |
O IV | ripe stage |
O V | spent stage |
OG | oogonium |
OC | oocyte |
N | nucleus |
CM | cytoplasmic membrane |
Y | yolk granule |
FC | follicle cell |
FM | follicle membrane |
CT | collecting tissue |
SG | spermatogonium |
SC | spermatocyte |
SC1 | primary spermatocyte |
SC2 | second spermatocyte |
ST | spermatid |
S | sperm |
WE | wall epithelium |
EM | eosinophilic matrix |
VD | vas deferens |
EB | the ejaculatory bulb |
ORF | open reading frame |
M. nipponense | Macrobrachium nipponense |
M. rosenbergii | Macrobrachium rosenbergii |
cDNA | complementary DNA |
FFRC | freshwater Fisheries Research Centre |
ISH | in situ hybridization |
CISH | chromogenic in situ hybridization |
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Primer Name | Sequence (5′-3′) | Description |
---|---|---|
DMRTD-F1 | TGCGCCMGRTGYMGRAAYCAYGG | For DM-domain RACE |
DMRTD-R1 | ARSGCSACYTGSGCSGCCATNAC | For DM-domain RACE |
DMRTD-R2 | TGSGCSGCCATNACCCKYTGCC | For DM-domain RACE |
5′-RACE outer | ATTACCCGTTGCCTCT | For 5′-RACE |
5′-RACE inner | ACCACCAGCAGGCAGTTG | For 5′-RACE |
DMRT-5′R | GCTTCTTGTGCCCCTTCA | For 5′-RACE |
3′-RACE outer | GGTGGTGTCCTGCCTGAA | For 3′-RACE |
3′-RACE inner | CCTGAAGGGGCACAAGAA | For 3′-RACE |
DMRT-3′F | CCTGAAGGGGCACAAGAA | For 3′-RACE |
DMRT-qF | ACGACCTTAGTAGGATGGACAGT | For RT-PCR |
DMRT-qR | GAGTGGAGGCAATAGAATGGGTA | For RT-PCR |
β-actinF | TATGCACTTCCTCATGCCATC | For RT-PCR |
β-actinR | AGGAGGCGGCAGTGGTCAT | For RT-PCR |
DMRT-P | CACTCCAGAATACTCGCCGAAGTCCTACGCGAAGAC | Probe foe ISH |
DMRT-iF | TAATACGACTCACTATAGGGCTTAGTGTCAGACGCGACCA | For DMRT dsRNA |
DMRT-iR | TAATACGACTCACTATAGGGCTTCGGCGAGTATTCTGGAG | For DMRT dsRNA |
IAG-qF | CGCCTCCGTCTGCCTGAGATAC | For RT-PCR |
IAG-qR | CCTCCTCCTCCACCTTCAATGC | For RT-PCR |
VG-qF | GAAGTTAGCGGAGATCTGAGGT | For RT-PCR |
VG-qR | CCTCGTTGACCAATCTTGAGAG | For RT-PCR |
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Wang, Y.; Jin, S.; Fu, H.; Qiao, H.; Sun, S.; Zhang, W.; Jiang, S.; Gong, Y.; Xiong, Y.; Wu, Y. Identification and Characterization of the DMRT11E Gene in the Oriental River Prawn Macrobrachium nipponense. Int. J. Mol. Sci. 2019, 20, 1734. https://doi.org/10.3390/ijms20071734
Wang Y, Jin S, Fu H, Qiao H, Sun S, Zhang W, Jiang S, Gong Y, Xiong Y, Wu Y. Identification and Characterization of the DMRT11E Gene in the Oriental River Prawn Macrobrachium nipponense. International Journal of Molecular Sciences. 2019; 20(7):1734. https://doi.org/10.3390/ijms20071734
Chicago/Turabian StyleWang, Yabing, Shubo Jin, Hongtuo Fu, Hui Qiao, Shengming Sun, Wenyi Zhang, Sufei Jiang, Yongsheng Gong, Yiwei Xiong, and Yan Wu. 2019. "Identification and Characterization of the DMRT11E Gene in the Oriental River Prawn Macrobrachium nipponense" International Journal of Molecular Sciences 20, no. 7: 1734. https://doi.org/10.3390/ijms20071734