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The functions and mechanisms of sequence differences of DGAT1 gene on milk fat synthesis between dairy cow and buffalo
Journal of Dairy Research ( IF 2.1 ) Pub Date : 2020-06-02 , DOI: 10.1017/s0022029920000126 Dinesh Bhattarai 1 , Rahim Dad 1 , Tesfay Worku 1 , Sutong Xu 1 , Farman Ullah 1 , Min Zhang 1 , Xianwei Liang 2 , Tingxian Den 2 , Mingxia Fan 3 , Shujun Zhang 1
Journal of Dairy Research ( IF 2.1 ) Pub Date : 2020-06-02 , DOI: 10.1017/s0022029920000126 Dinesh Bhattarai 1 , Rahim Dad 1 , Tesfay Worku 1 , Sutong Xu 1 , Farman Ullah 1 , Min Zhang 1 , Xianwei Liang 2 , Tingxian Den 2 , Mingxia Fan 3 , Shujun Zhang 1
Affiliation
In this research communication we describe the DGAT1 sequence and promoter region in dairy cows and buffalo and compare the activities of DGAT1 between the two species in order to increase knowledge of the cause of milk fat variation. pGL-3 basic vectors were used to construct the reporter gene. Based on the predicted promoter region, 4 truncated plasmid vectors were constructed in cow-DGAT1 and 3 plasmid vectors in buffalo-DGAT1. Each reporter plasmid was transfected into the bovine mammary epithelial cell (BMEC), 293T cell, and CHO cells to analyze the activity using Dual-Luciferase Reporter Assay System. The results show that the region between −93 to −556 bp was essential for cow promoter activity while −84 to −590 bp was essential for buffalo promoter activity revealing these regions contain core promoter. The buffalo has higher promoter activity than cow yet it was not statistically significant. Comparison of candidate mutation K232A between cow and buffalo population revealed the presence of both the allelic population in dairy cows (lysine and alanine) however, only K (lysine) allelic amino acid was found in buffalo population. The absence of the alanine allelic population from buffalo explains the higher fat content of buffalo milk.
中文翻译:
DGAT1基因序列差异对奶牛和水牛乳脂合成的作用及机制
在本研究交流中,我们描述了奶牛和水牛的 DGAT1 序列和启动子区域,并比较了两个物种之间 DGAT1 的活性,以增加对乳脂变异原因的认识。pGL-3基本载体用于构建报告基因。基于预测的启动子区域,在cow-DGAT1中构建了4个截短的质粒载体,在buffalo-DGAT1中构建了3个质粒载体。将每个报告质粒转染到牛乳腺上皮细胞 (BMEC)、293T 细胞和 CHO 细胞中,以使用 Dual-Luciferase Reporter Assay System 分析活性。结果表明,-93 至 -556 bp 之间的区域对奶牛启动子活性至关重要,而 -84 至 -590 bp 对水牛启动子活性至关重要,这表明这些区域包含核心启动子。水牛的启动子活性比奶牛高,但没有统计学意义。牛和水牛种群之间的候选突变 K232A 的比较揭示了奶牛中存在等位基因种群(赖氨酸和丙氨酸),然而,在水牛种群中仅发现 K(赖氨酸)等位基因氨基酸。来自水牛的丙氨酸等位基因群的缺失解释了水牛奶中较高的脂肪含量。
更新日期:2020-06-02
中文翻译:
DGAT1基因序列差异对奶牛和水牛乳脂合成的作用及机制
在本研究交流中,我们描述了奶牛和水牛的 DGAT1 序列和启动子区域,并比较了两个物种之间 DGAT1 的活性,以增加对乳脂变异原因的认识。pGL-3基本载体用于构建报告基因。基于预测的启动子区域,在cow-DGAT1中构建了4个截短的质粒载体,在buffalo-DGAT1中构建了3个质粒载体。将每个报告质粒转染到牛乳腺上皮细胞 (BMEC)、293T 细胞和 CHO 细胞中,以使用 Dual-Luciferase Reporter Assay System 分析活性。结果表明,-93 至 -556 bp 之间的区域对奶牛启动子活性至关重要,而 -84 至 -590 bp 对水牛启动子活性至关重要,这表明这些区域包含核心启动子。水牛的启动子活性比奶牛高,但没有统计学意义。牛和水牛种群之间的候选突变 K232A 的比较揭示了奶牛中存在等位基因种群(赖氨酸和丙氨酸),然而,在水牛种群中仅发现 K(赖氨酸)等位基因氨基酸。来自水牛的丙氨酸等位基因群的缺失解释了水牛奶中较高的脂肪含量。