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Identification of the Core Promoter Region of KCNJ15 Gene in Chengde Polled Goat (Capra hircus) |
ZHANG Wen-Hao, LIU Shuang, HAN Cong, HONG Zhi-Lin, XIE Yu-Chun, QIAO Xian, LIU Zheng-Zhu*, GONG Yuan-Fang* |
College of Animal Science and Technology, Hebei Normal University of Science and Technology/Hebei Key Laboratory of Specially Animal Germplasm Resources Exploration and Innovation, Qinhuangdao 066004, China |
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Abstract Horn is an important trait in goats (Capra hircus), and the potassium inwardly rectifying channel subfamily J member 15 (KCNJ15) gene is an important gene, which affects the growth of horn. The present study selected Chengde polled goat as the test animal and extracted genomic DNA. Then the 2 353 bp sequence (2064 bp upstream~289 bp downstream of the start codon) of KCNJ15 gene of Chengde polled goat was amplified by PCR. The sequence was analyzed by online software, the plasmids with different deletion fragments were constructed and transiently transfected into 293T cells. In order to determine the core promoter region of the KCNJ15 gene, dual luciferase detection kit was used to measure the activity. The sequence of Chengde polled goat (2353 bp) was successfully obtained, and software prediction showed that -423~+289 bp might be the active region in the promoter. At the same time, several transcription factor binding sites and CAAT and TATA regulatory elements were simultaneously predicted. The dual luciferase activity assay showed that -491~+ 289 bp had the highest relative activity value among the 4 fragments, suggesting that this region might be the core promoter region, which was consistent with the results predicted by software. This study provides basic data for in-depth exploration of the molecular regulation mechanism of KCNJ15 gene in goat horn traits.
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Received: 21 August 2023
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Corresponding Authors:
* liuzhengzhu@163.com; gyfkeyan@163.com
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