中国医科大学学报

中国医科大学学报

中国医科大学学报 ›› 2009, Vol. 38 ›› Issue (9): 647–.

• 基础医学 •    下一篇

人巨细胞病毒UL135蛋白结合蛋白的酵母双杂交筛选

  

  • 出版日期:2009-09-20 发布日期:2010-03-30
  • 通讯作者: 孙峥嵘 E-mail:sunzr@sj hospital.org

Screening of Protein Interacting with HCMV UL135 Protein by Yeast Two Hybrid System from Human Fetus Brain cDNA Library

  • Online:2009-09-20 Published:2010-03-30
  • Contact: SUN Zheng-Rong E-mail:sunzr@sj hospital.org

摘要:

目的利用酵母双杂交系统从人胎脑cDNA文库中筛选与人巨细胞病毒(HCMV)UL135编码蛋白相互作用的蛋白。方法将成功构建的酵母诱饵表达载体pGBKT7-UL135转化到酵母菌AH109中,再将人胎脑文库DNA转化到含有pGBKT7-UL135的酵母细胞中,筛选与HCMVUL135编码蛋白相互作用的细胞蛋白,并对阳性克隆进行测序和生物信息学分析。结果最终确认60个克隆与HCMVUL135编码蛋白相互作用,其中2个克隆与Thy-1高度同源。结论成功应用酵母双杂交系统筛选出与HCMVUL135编码蛋白相互作用的蛋白,其中Thy-1在HCMV感染致病过程中可能起重要作用。

关键词: 人巨细胞病毒, UL135蛋白, 酵母双杂交系统, 人胎脑cDNA文库, 人巨细胞病毒, UL135 蛋白, 酵母双杂交系统, 人胎脑cDNA文库

Abstract:

bjective To screen the proteins which can interact with the human
      cytomeg a lovirus HCMV UL135 protein from human fetus brain cDNA library
      with yeast two-hybrid system. Methods The "bait plasmid"(named as
      pGBKT7-UL135)was constructed. Using HCMV UL135 as a bait,we screened a
      human fetus brain cDNA library. The proteins interacting with UL135
      protein were searched. The positive clones were sequenced and analyzed by
      bioinformatic methods. Results Ten clones interacting with HCMV UL135 were
      identified, two of them were Thy-1. Conclusion Some proteins interacting
      with HCMV UL135 in human fetus brain cDNA library were successfully
      screened. Thy-1 may play an important role in the course of HCMV
infection.

Key words: human cytomegalovirus, UL135 protein, yeast two-hybrid system; human fetus brain cDNA library, HCMV, UL135 protein, Two-Hybrid System, Human fetus brain cDNA Library

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[3] 柳中洋,齐莹,郭鑫,蒋树娟,黄郁晶,邵耀中,阮强. 人巨细胞病毒微小RNA miR?UL70?5P靶向抑制人即刻早期反应蛋白3的表达[J]. 中国医科大学学报, 2014, 43(7): 577-580.
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[5] 卢志涛,王桂丽,马艳萍,崔鑫,任高伟,齐莹,阮强,孙峥嵘. 人巨细胞病毒UL141编码蛋白相互作用蛋白的筛选和分析[J]. 中国医科大学学报, 2013, 42(10): 865-868.
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