| 田侠 徐东刚 杜建芳.BAFF-R 胞内区与TRAF3 相结合关键分子结合域的确定[J].,2015,15(8):1444-1448 |
| BAFF-R 胞内区与TRAF3 相结合关键分子结合域的确定 |
| Identification of the Key Molecular Binding Domains of the CytoplasmicDomain of BAFF-R Binding with TRAF3 |
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| DOI： |
| 中文关键词: BAFF-R TRAF3 突变体 |
| 英文关键词: BAFF-R TRAF3 Mutants |
| 基金项目:国家自然科学基金项目（30901795） |
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| 中文摘要: |
| 目的：研究BAFF-R 胞内区与TRAF3 相互作用的关键结合域。方法：分别钓取BAFF-R 胞内区和TRAF3 的cDNA 片段，克
隆到酵母双杂交系统中，验证两者之间的相互作用。利用缺失PCR和重叠PCR 的方法获取11 个TRAF3 的突变体，验证TRAF3
的11 个突变体和BAFF-R 胞内区的相互作用。结果：TRAF3 与BAFF-R胞内区发生相互作用时，有三个关键的分子结合域，分别
是N 端的螺旋结构382- 400 位氨基酸、中间的428－463 位氨基酸以及TRAF3 C端的543－560 位氨基酸。结论：在体内得到了
BAFF-R胞内区与TRAF3 相互作用的三个关键分子结合域，有助于展开对BAFF-R 胞内区与TRAF3 相互作用的进一步研究。 |
| 英文摘要: |
| Objective:To identify the key cytoplasmic molecular domain of BAFF-R binding with TNFR-associated factor 3
(TRAF3).Methods:At first, we gained the cDNA of cytoplasmic domain of BAFF-R and TRAF3 respectively by RT-PCR. Then, we
used yeast two-hybrid assay to testify the interactions between the cytoplasmic domain of BAFF-R with TRAF3. In order to gain the key
molecular binding domains of BAFF-R with TRAF3, the eleven deletion mutants were constructed by PCR and overlapped PCR. At last,
we used yeast two-hybrid assay to testify the interaction between the eleven mutants with TRAF3.Results:382-400aa coil, 428-463aa,
543-560aa of TRAF3 were three important domains which play critical part in binding to BAFF-R.Conclusion:The result will provide a
molecular binding base for the association of cytoplasmic domain of BAFF-R with TRAF3. |
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