目的探讨乙型肝炎病毒(HBV)HBx蛋白(HepatitisB virus X protein)对α干扰素(IFN-α)诱导的抗病毒蛋白的影响及相关机制。方法以表达HBx蛋白的重组质粒FL1-145HBx转染人肝胚瘤细胞株HepG2细胞,经IFN-α处理后,RT-PCR法分析细胞内抗病毒...目的探讨乙型肝炎病毒(HBV)HBx蛋白(HepatitisB virus X protein)对α干扰素(IFN-α)诱导的抗病毒蛋白的影响及相关机制。方法以表达HBx蛋白的重组质粒FL1-145HBx转染人肝胚瘤细胞株HepG2细胞,经IFN-α处理后,RT-PCR法分析细胞内抗病毒蛋白MxA和JAK-STAT信号转导途径分子STAT1 mRNA表达水平,同时运用免疫印迹检测细胞内HBx、p-ERK、p-STAT1和t-STAT1等蛋白的表达。结果转染细胞内MxA、STAT1的mRNA和p-STAT1、t-STAT1的蛋白表达水平明显减少(P<0.05);而ERK抑制剂PD98059预处理后,转染细胞内MxA、STAT1 mRNA水平能够恢复至转染前的表达水平。结论 HBx蛋白很可能通过影响IFN-αJAK-STAT信号转导途径分子而抑制抗病毒蛋白MxA的表达;ERK信号转导途径的活化可能参与这一抑制过程。展开更多
[Objective] The study aimed to establish a fast and accurate method to detect the polymorphism of the 12^th exon of equine MxA gene. [Method] The 12^th exon of MxA gene was amplified by mismatch PCR and the products w...[Objective] The study aimed to establish a fast and accurate method to detect the polymorphism of the 12^th exon of equine MxA gene. [Method] The 12^th exon of MxA gene was amplified by mismatch PCR and the products were analyzed by restriction fragment length polymorphism （RFLP） to determine the point mutation at the 1 790 nt of MxA cDNA. The sequence of the PCR products was also analyzed. [Result] There were three genotypes （AA, AB and BB） in the 12^th exon of equine MxA gene; the 2 081 nt of MxA cDNA mutated from G to C, correspondingly changing the 562^th amino acid of the coding region of MxA protein from tryptophan to cysteine; the specific sequence of the PCR products amplified by mismatch PCR-RFLP was consistent with the analysis results of RFLP. [ Conclusion] The mismatch PCR-RFLP was an easy method with accurate results to detect the polymorphism of the 12^th exon of equine MxA gene.展开更多
基金Supported by the Natural Science Foundation of Inner Mongolia Au-tonomous Region (200508010413)~~
文摘[Objective] The study aimed to establish a fast and accurate method to detect the polymorphism of the 12^th exon of equine MxA gene. [Method] The 12^th exon of MxA gene was amplified by mismatch PCR and the products were analyzed by restriction fragment length polymorphism （RFLP） to determine the point mutation at the 1 790 nt of MxA cDNA. The sequence of the PCR products was also analyzed. [Result] There were three genotypes （AA, AB and BB） in the 12^th exon of equine MxA gene; the 2 081 nt of MxA cDNA mutated from G to C, correspondingly changing the 562^th amino acid of the coding region of MxA protein from tryptophan to cysteine; the specific sequence of the PCR products amplified by mismatch PCR-RFLP was consistent with the analysis results of RFLP. [ Conclusion] The mismatch PCR-RFLP was an easy method with accurate results to detect the polymorphism of the 12^th exon of equine MxA gene.
