Characterization of the Expression Profile and Genetic Polymorphism of the Cellular Retinol-Binding Protein (CRBP IV) Gene in Erlang Mountainous Chickens
Abstract
:1. Introduction
2. Results
2.1. Molecular Cloning and Sequence of the Chicken CRBP IV Gene
2.2. Ontogenic Expression of CRBP IV Gene in Chickens
2.3. Comparison of CRBP IV mRNA Expression among Different Tissues in Chickens
2.4. Identification of Genetic Variants in Chicken CRBP IV Gene
2.5. Genotype Frequencies and Association Analysis with Egg Production Traits
3. Discussion
4. Experimental Section
4.1. Chicken Populations and Trait Measurement
4.2. Total DNA Extraction, RNA Isolation and Reverse Transcription
4.3. Cloning and Sequencing of PCR Product
4.4. Real-Time PCR, SNPs Identification and Association Analysis
4.5. Data Analysis
5. Conclusion
Acknowledgments
References
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Species | GenBank accession number | Amino acid Identity 1 |
---|---|---|
Gallus gallus | XP_417606.2 | 98.4% (125/127) |
Sus scrofa | NP_001138694.1 | 74.8% (95/127) |
Mus musculus | NP_071303.1 | 72.4% (92/127) |
Rattus norveicus | NP_001102163 | 71.7% (91/127) |
Homo sapiens | NP_443192.1 | 70.9% (90/127) |
Pan troglodytes | XP_001161299.1 | 70.9% (90/127) |
Bos taurus | XP_871364.3 | 66.9% (85/127) |
Canis lupus familiaris | XP_536740.1 | 57.5% (73/127) |
Lines | N. | Alleles frequency | Genotypes frequency | p values of chi-square 1 | PIC2 | |||
---|---|---|---|---|---|---|---|---|
T | C | TT | TC | CC | ||||
SD02 | 188 | 0.641 | 0.359 | 0.399 | 0.484 | 0.117 | 0.471 | 0.354 |
SD03 | 161 | 0.630 | 0.370 | 0.373 | 0.515 | 0.112 | 0.182 | 0.357 |
Total | 349 | 0.636 | 0.364 | 0.386 | 0.499 | 0.115 | 0.091 | 0.356 |
Items | N | Traits | ||||||
---|---|---|---|---|---|---|---|---|
AFE | BWFE | FEW | TE300 | HCLD | ALI | |||
Genotypes | CC | 40 | 169.3 ± 10.0 b | 1981.5 ± 296.5 | 40.4 ± 4.9 | 85.6 ± 20.4 a,A | 9.7 ± 3.8 a | 1.8 ± 1.2 |
TC | 174 | 175.1 ± 11.2 a | 1892.1 ± 244.5 | 39.9 ± 5.8 | 81.1 ± 16.9 b,A,B | 7.8 ± 3.6 b | 1.6 ± 0.7 | |
TT | 135 | 176.5 ± 11.5 a | 1898.0 ± 247.9 | 40.5 ± 6.8 | 77.9 ± 18.2 c,B | 7.2 ± 3.2 b | 1.7 ± 0.8 | |
Lines | SD02 | 188 | 169.8 ± 10.2 b | 1903.0 ± 241.2 | 40.2 ± 6.6 | 83.3 ± 18.4 | 7.7 ± 4.0 | 1.6 ± 0.9 |
SD03 | 161 | 176.8 ± 10.9 a | 1919.5 ± 250.2 | 41.3 ± 5.3 | 81.3 ± 17.3 | 7.8 ± 4.5 | 1.5 ± 0.8 |
Primers | Primer Sequence (5′→3′) | Annealing Temperature (°C) | Product Length (bp) | Amplified Region |
---|---|---|---|---|
Primer pairs for measuring cloning and sequencing chicken CRBP IV gene | ||||
CRBPIV-F | TGACTGTAACTCCTTCCCACAGTCT | 56 | 464 | 3–466 |
CRBPIV-R | GAAGACAGCTTGCACCTTATGAC | |||
Primer pairs for measuring chicken CRBP IV gene expression | ||||
CRBPIV-F | CATACCACAAGCACATTCAGAGA | 58 | 125 | 1320–1444 |
CRBPIV-R | AGTTTGTCATTGTCCCAGGTAAC | |||
β-actin-F | GAGAAATTGTGCGTGACATCA | 60 | 152 | 685–836 |
β-actin-R | CCTGAACCTCTCATTGCCA | |||
Primer pairs for screening chicken CRBP IV gene polymorphisms | ||||
P1-F | CTTCCCACAGTCTAGCAA | 56.6 | 236 | 3380706–3380919 |
P1-R | ATCAGCTAACAAATATCCTTC | |||
P2-F | TTCTTCTGTCAAAGGTATTG | 55.2 | 244 | 3381431–3381654 |
P2-R | CTTCCTTCTGTAAGAACACAT | |||
P3-F | CAGAGCCTGGTTACCTG | 56.6 | 180 | 3381893–3382052 |
P3-R | AGCTTGCACCTTATGACTACA |
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Yin, H.-D.; Wang, Y.; Zhang, Z.-C.; Liu, Y.-P.; Chen, S.-Y.; Zhu, Q. Characterization of the Expression Profile and Genetic Polymorphism of the Cellular Retinol-Binding Protein (CRBP IV) Gene in Erlang Mountainous Chickens. Int. J. Mol. Sci. 2013, 14, 4432-4443. https://doi.org/10.3390/ijms14034432
Yin H-D, Wang Y, Zhang Z-C, Liu Y-P, Chen S-Y, Zhu Q. Characterization of the Expression Profile and Genetic Polymorphism of the Cellular Retinol-Binding Protein (CRBP IV) Gene in Erlang Mountainous Chickens. International Journal of Molecular Sciences. 2013; 14(3):4432-4443. https://doi.org/10.3390/ijms14034432
Chicago/Turabian StyleYin, Hua-Dong, Yan Wang, Zhi-Chao Zhang, Yi-Ping Liu, Shi-Yi Chen, and Qing Zhu. 2013. "Characterization of the Expression Profile and Genetic Polymorphism of the Cellular Retinol-Binding Protein (CRBP IV) Gene in Erlang Mountainous Chickens" International Journal of Molecular Sciences 14, no. 3: 4432-4443. https://doi.org/10.3390/ijms14034432