摘要
为了探讨NLRC5启动子的潜在调控机制,将NLRC5基因启动子系列缺失片段插入到pGL3-basic载体,构建重组质粒,并转染DF1细胞系。通过双荧光素酶实验寻找核心调控区,然后用目标捕获测序检测27个鸡品种的NLRC5核心启动子区的SNPs,并利用TRANSFAC,JASPAR和Meth Primer预测该区域的转录因子结合位点和CpG岛。结果显示,NLRC5启动子有2个核心区域,分别是1—617和1448—2108。第一个核心区域内存在3个SNPs,其中SNP1影响转录因子Hic1的结合序列,但SNPs对CpG岛不产生影响,表明NLRC5基因的启动子区的调控可能受不同因素的影响,但SNPs并不在甲基化的水平上影响启动子的活性。
In order to explore the potential regulation mechanism of chicken NLRC5 promoter,deletion fragments of chicken NLRC5 promoter was cloned and inserted into pGL3-basic vector to construct recombinant plasmids for transfecting DF1 cells. Dual-luciferase assay was carried out to find the core regulation region. Then target sequence capture assay was used to detect SNPs in the core region of NLRC5 promoter of 27 chicken breeds. Finally,transcription factor binding sites and CpG islands were predicted by using online softwares TRANSFAC,JASPAR and Meth Primer. The results showed that NLRC5 promoter had 2 core regions,1 to 617 and 1 448 to 2 108,respectively. There were 3 SNPs loci totally in the first core region in all breeds. SNP1 was found to influence the transcription factor Hic1 binding site. However,SNPs in the first core region did not affect CpG island. The results above suggested that NLRC5 promoter activity might be regulated by different factors,but SNPs didn＇t influence the promoter activity at methylation level.
出处
《浙江农业学报》
CSCD
北大核心
2017年第3期395-400,共6页
Acta Agriculturae Zhejiangensis
基金
国家科技支撑计划(2015BAD03B03)
江苏省六大人才高峰项目(2015-NY-019)
