为揭示牛MHC-DRB3上游调控区多态对其表达调控作用,利用PCR-SSCP和DNA序列测定技术分析并建立BoLA-DRB3*exon2和BoLA-DRB3 URR之间的连锁关系。结合不同近端调控区与转录因子结合模式预测结果,选择3种不同上游调控序列的连锁组合,应用实时定量RT-PCR技术对牛MHC-DRB3基因mRNA相对表达量进行了验证分析。结果表明,上游调控区基因型为H17H17的BoLA-DRB3基因mRNA相对表达量显著高于(P<0.01)基因型H1H1及H5H6。初步证明上游调控区中的多态会引起结构基因表达量的变化。
王学清
,
李魁英
,
裴翠娟
,
张峰
,
吴占军
,
刘小虎
,
马书林
,
张新同
,
王昆
. 中国荷斯坦牛BoLA-DRB3基因的mRNA表达研究[J]. 华北农学报, 2013
, 28(4)
: 19
-22
.
DOI: 10.3969/j.issn.1000-7091.2013.04.004
To study the regulation of upstream regulatory region polymorphisms on BoLA- DRB3 gene expres- sion,PCR- SSCP and DNA sequencing were used to identify the linkage relationship between BoLA- DRB3* exon2 and BoLA- DRB3 URR.Combined with the predicted results of binding mode between different URR and transcrip- tion factor,three linkage types were selected to detect the BoLA-DRB3 mRNA levels by relative Real- time PCR ex- periments. The results showed that the expression abundances of BoLA-DRB3 varied in different linkage types.The expression level of mRNA of BoLA- DRB3 gene linked with H17H17 was significant higher than H1H1 and H5H6 (P<0.01).This preliminary proved that the polymorphism of upstream regulatory region can cause the expression change of BoLA- DRB3 structural gene.
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