APOBEC3B羧基端抗乙肝病毒位点的筛选和鉴定

发布时间：2018-05-08 12:52

本文选题：APOBECB + 脱氨基　；参考：《重庆医科大学学报》2017年07期

【摘要】：目的:筛选参与APOBEC3B羧基端脱氨酶及抗乙肝病毒(hepatitis B virus,HBV)活性的重要区域和关键位点。方法:定点突变构建羧基端活性中心附近的3个螺旋区段(α2、α3、α4)的缺失突变体和α4螺旋区内(D316、K320、E321、R327、D328)位点的突变体;Southern blot筛选APOBEC3B抑制HBV复制的重要区段和关键位点;不同DNA变性PCR(differential DNA denaturation PCR,3D-PCR)和克隆测序进行验证。结果:APOBEC3B羧基端脱氨酶活性中心周围的α3和α4螺旋区缺失后,其抗病毒活性消失;D316位点突变后APOBEC3B的脱氨酶活性和抗病毒活性消失。结论:APOBEC3B羧基端的D316位点是APOBEC3B脱氨酶和抗病毒的关键位点。
[Abstract]:Aim: to screen the important regions and key sites involved in the activities of APOBEC3B carboxyl terminal deaminase (APOBEC3B) and anti hepatitis B virus (HBV). Methods: three deletion mutants (伪 _ 2, 伪 _ 3, 伪 _ 4) were constructed by site-directed mutagenesis. Southern blot was used to screen the important regions and key sites of APOBEC3B for inhibiting HBV replication. Different DNA denaturation PCR(differential DNA denaturation PCR3 D-PCR) and clone sequencing were verified. Results the antiviral activity of APOBEC3B disappeared after the deletion of 伪 3 and 伪 4 helical region around the carboxyl terminal deaminase activity center of 1: APOBEC3B. The deaminase activity and antiviral activity of APOBEC3B disappeared after mutation at D316 site. Conclusion the D316 site of the carboxyl terminal of APOBEC3B is the key site of APOBEC3B deaminase and antivirus.
【作者单位】：重庆医科大学感染性疾病分子生物学教育部重点实验室;重庆医科大学基础医学院病原微生物教研室;
【基金】：国家自然科学基金资助项目(编号:81471945) 重庆市科委自然科学基金资助项目(编号:cstc2014jcyj A10075) 重庆市教委科学技术研究资助项目(编号:KJ1600205)
【分类号】：R373

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