Characterization of GLOD4 in Leydig Cells of Tibetan Sheep during Different Stages of Maturity
Abstract
:1. Introduction
2. Materials and Methods
2.1. Experimental Animals and Design
2.2. Total RNA Extraction and cDNA Synthesis
2.3. Cloning of Sheep GLOD4 Gene
2.4. Bioinformatics Analysis of the GLOD4 Gene
2.5. qRT-PCR
2.6. Western Blot
2.7. Immunohistochemistry and Immunofluorescence
2.8. Image Analysis and Data Statistics
3. Results
3.1. CDS Sequence Characterization of GLOD4
3.2. Homology Analysis and Evolutionary Relationships of the GLOD4 Gene among Different Species
3.3. Expression Patterns of the GLOD4 Transcript at Different Developmental Stages of Tibetan Sheep Testes
3.4. Localization of the GLOD4 Protein in Developmental Sheep Testes
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
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Gene | GenBank No. | Sequence(5′-3′) | Length/bp | Utilization |
---|---|---|---|---|
GLOD4 | XM_027975163.1 | F: ATGGTGGGGTATGGACCCG | 729 | Cloning |
R: TTAACCTGTCGCCTTTGGTTTC | ||||
GLOD4 | XM_027975163.1 | F: CTCCGTTGGTGAGTCTGG | 167 | qRT- PCR |
R: ATCTCCTGCAATTGCGTCG | ||||
β-actin | NM_001009784.1 | F: CTTCCAGCCTTCCTTCCTGG | 180 | qRT- PCR |
R: GCCAGGGCAGTGATCTCTTT |
Species | Genbank No. | Nucleotide Similarity % | Amino acid Similarity % |
---|---|---|---|
Pantholops hodgsonii (chiru) | XM_005975769.1 | 99.18 | 100 |
Capra hircus (goat) | XM_018064369.1 | 99.18 | 100 |
Bos Taurus (cattle) | NM_001101953.1 | 96.57 | 97.52 |
Equus caballus (horse) | XM_001504263.5 | 87.79 | 85.95 |
Sus scrofa (pig) | XM_003131815.5 | 87.52 | 85.54 |
Camelus bactrianus (Bactrian camel) | XM_010958222.1 | 87.24 | 84.71 |
Felis catus (domestic cat) | XM_003996448.4 | 86.15 | 85.95 |
Canis lupus dingo (dingo) | XM_025476209.1 | 85.95 | 83.88 |
Homo sapiens (human) | XM_024450778.1 | 85.23 | 84.3 |
Mus musculus (house mouse) | NM_001362901.1 | 84.72 | 83.47 |
Gallus gallus (chicken) | NM_001282341.1 | 74.54 | 76.76 |
Mastacembelus armatus (zig-zag eel) | XM_026299138.1 | 69.74 | 68.18 |
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Wang, X.; Li, T.; Liu, N.; Zhang, H.; Zhao, X.; Ma, Y. Characterization of GLOD4 in Leydig Cells of Tibetan Sheep during Different Stages of Maturity. Genes 2019, 10, 796. https://doi.org/10.3390/genes10100796
Wang X, Li T, Liu N, Zhang H, Zhao X, Ma Y. Characterization of GLOD4 in Leydig Cells of Tibetan Sheep during Different Stages of Maturity. Genes. 2019; 10(10):796. https://doi.org/10.3390/genes10100796
Chicago/Turabian StyleWang, Xia, Taotao Li, Ningbo Liu, Hongyu Zhang, Xingxu Zhao, and Youji Ma. 2019. "Characterization of GLOD4 in Leydig Cells of Tibetan Sheep during Different Stages of Maturity" Genes 10, no. 10: 796. https://doi.org/10.3390/genes10100796