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Screening and identification of interacting proteins with hepatitis B virus core protein in leukocytes and cloning of new gene C1
  • ISSN号:1007-9327
  • 期刊名称:《世界胃肠病学杂志:英文版》
  • 时间:0
  • 分类:R512.62[医药卫生—临床医学;医药卫生—内科学]
  • 作者机构:[1]Department of Infectious Diseases, The First Affiliated Hospital of Medical College of Xi'an Jiaotong University, Xi'an 710061, Shaanxi Province, China, [2]Institute of Infectious Diseases, Ditan Hospital, 13 Ditan Park, Anwai Street, Dongcheng District, Beijing 100011, China
  • 相关基金:Supported by the National Natural Science Foundation of China, No. 30471532
中文摘要:

瞄准:在外部血在 HBV 复制的致病调查 HBcAg 的生物功能单音的原子房间(PBMC ) 。方法:HBcAg 区域被聚合酶链反应(PCR ) 放大, HBV HBcAg 诱饵原生质标志 pGBKT7-HBcAg 被平淡的分子的生物方法构造。然后, recombinant 原生质标志 DNA 被转变成酵母 AH109。在 HBV 核心蛋白质在 AH109 酵母紧张(西方的污点分析) 被表示以后,酵母 -- 屏蔽的二个混血儿被与包含白血球 cDNA 图书馆的 Y187 交配 AH109 执行原生质标志。双酵母房间是合成退学学生营养培养基(SD/-Trp-Leu-His-Ade )(QDO ) 和合成退学学生营养培养基(SD/-Trp-Leu-His-Ade )(TDO ) 上的 plated。第二屏蔽与 LacZ 报告基因被执行(酵母房间在 QDO 被种中等包含 X-alpha-gal ) 。在从积极殖民地获得的 HBV 核心蛋白质和蛋白质之间的相互作用被重复酵母进一步证实 -- 二个混血儿。在原生质标志 DNA 从蓝殖民地被提取并且定序以后,结果被生物信息的方法分析。结果:十八个殖民地被获得并且定序,包括在癌症 2 的亢奋的甲基化( 3 关口),真核细胞的翻译延伸因素 2 ( 2 关口),乙酰辅酶 A 合成酶 3 ( 1 个关口一个), DNA 聚合酶鲸鱼群妈( 1 个关口一个),通常认为的翻译开始因素( 1 个关口一个), chemokine ( C-C 主题)受体 5 ( 1 个关口一个), mitochondrial ribosomal 蛋白质 L41 ( 1 个关口一个), kyot 绑定蛋白质基因( 1 个关口一个), RanBPM ( 1 个关口一个),HBeAg有约束力的蛋白质 3 ( 1 个关口一个),规划了细胞死亡 2 ( 1 个关口一个)。有未知功能的四新基因被识别。结论:在白血球的核心蛋白质交往蛋白质可以提供的 HBV 的基因的成功的克隆为学习 HBV 的生物功能的一些新线索核心蛋白质。

英文摘要:

AIM: To investigate the biological function of HBcAg in pathogenesis of HBV replication in peripheral blood mononuclear cells (PBMCs). METHODS: HBcAg region was amplified by polymerase chain reaction (PCR) and HBV HBcAg bait plasmid pGBKT7-HBcAg was constructed by routine molecular biological methods. Then the recombinant plasmid DNA was transformed into yeast AH109. After the HBV core protein was expressed in AH109 yeast strains (Western blot analysis), yeast-two hybrid screening was performed by mating AH109 with Y187 containing leukocyte cDNA library plasmid. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His- Ade) (QDO) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) (TDO). The second screening was performed with the LacZ report gene ( yeast cells were grown in QDO medium containing X-a-gal). The interaction between HBV core protein and the protein obtained from positive colonies was further confirmed by repeating yeast-two hybrid. After plasmid DNA was extracted from blue colonies and sequenced, the results were analyzed by bioinformatic methods. RESULTS: Eighteen colonies were obtained and sequenced, including hypermethylated in cancer 2 (3 colones), eukaryotic translation elongation factor 2 (2 colones), acetyl-coenzyme A synthetase 3 (1 colone), DNA polymerase gamma (1 colone), putative translation initiation factor (1 colone), chemokine (C-C motif) receptor 5 (1 colone), mitochondrial ribosomal protein L41 (1 colone), kyot binding protein genes (1 colone), RanBPM (1 colone), HBeAg-binding protein 3 (1 colone), programmed cell death 2 (1 colone). Four new genes with unknown function were identified. CONCLUSION: Successful cloning of genes of HBV core protein interacting proteins in leukocytes may provide some new clues for studying the biological functions of HBV core protein.

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  • 《世界胃肠病学杂志:英文版》
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  • 国际标准刊号:ISSN:1007-9327
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