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| Identification of the Core Promoter and SNPs Analysis of TYR Gene in Bashang Long-tail Chicken (Gallus gallus) |
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Abstract Abstract As a key enzyme involved in the formation and regulation of melanin, tyrosinase (TYR) has an important role in animal hair color phenotype. Tyrosinase dysfunction can block the pathway of melanin formation. The mutation of tyrosinase causes white phenotype of skin, hair, and feathers. Transcriptional regulation of gene promoter region plays an important role in gene expression. The research was designed to study the activity and the structure of TYR promoter. Dual-luciferase expression vectors were constructed and transfected to chicken embryo fibroblasts DF1 cells. The dual-luciferase detection kit was used to measure the relative luciferase activity. The length of the fragment in 5′ flanking region of TYR gene in chicken was 1 813 bp. Seven fragments with different length of promoter regions were amplified and cloned into the vector pGL3-Basic. Two mutant vectors of the promoter region were constructed. The core promoter region from -810 bp to +65 bp was identified in chicken TYR gene. The region from -810 bp to -213 bp may positively regulate TYR gene during the transcription process. The transcription factor (SP1, Oct-1, GATA-1, C/EBP, SRF, NF-1, YY1, NF-κB, TBP, and AP-1) may play a pivotal role in the gene transcription. The polymorphic sites of -963~-957, -955, -932, -692, -391, and -321 were found and the function of these sites in regulating promoter activity was critical but not all. Eight haplotypes (Hap1~Hap8) were predicted by the SHEsis software. The difference of genotype and allele frequency between white feather and black feather chicken was significant (P＜0.05). The dominant genotypes of white feather chicken were BB, DD, FF, II, KK, and MM respectively, the frequency of Hap1 was 100%. The dominant genotypes of black feather chicken were AA, CD, EF, HH, JJ, and LL respectively, the frequency of Hap2 had the highest proportion of haplotype (40%). The promoter activity value of mut-2 was significantly lower than P2, and the promoter activity value of mut-1 was significantly lower than P6 (P＜0.05). The decrease of promoter activity may affect TYR gene transcription, thus affecting the synthesis of melanin. The present study defines the activity and the structure of TYR promoter and provides the basis for studying the expression of this gene in hair follicle tissue.
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Received: 12 March 2018
Published: 21 May 2018
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ZHAO Zhi-Dong, TIAN Hong-Shan, JIANG Yan-Yan, SHI Bin-Gang, LIU Xiu, LI Xu-Peng, WANG Deng-Zhe, CHEN Jin-Lin, HU Jiang. Polymorphisms of ACSL1 Gene Promoter and Their Association Analysis with Milk Quality Traits in Yak (Bos grunniens)[J]. 农业生物技术学报, 2019, 27(9): 1596-1603. |
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