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Toll样受体2胞外域及其氨基端和羧基端片段的克隆与序列分析 被引量：2

Cloning and sequencing of extracellular domain and N-Terminal and C-Terminal fragments of Toll-like receptor 2
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摘要 目的　为研究 TL R2胞外域在肽聚糖诱导的信号转导中的功能 ,构建 TL R2胞外域及其氨基端和羧基端片段真核表达载体 .方法　提取 HL - 6 0细胞总 RNA,以 RT- PCR方法获取了 TL R2胞外域 c DNA片段 ,将其与 p GEM- T Easy载体连接 ,转化 E.coli JM10 9,建立了 TL R2胞外域的 c DNA克隆 ;籍此 ,又相继克隆了胞外域的氨基端和羧基端片段 ,然后将这三个片段克隆到真核表达载体 pc DNA3中 .结果　序列分析表明 ,与 Gen Bank中的人 TL R2全长 c DNA序列比较 ,仅 4个碱基不同 ,同源性为 0 .998.结论　成功获得了HL- 6 0细胞 TL AIM To find out the role of the extracellular domain of TLR2 in the peptidoglycan induced signaling pathway, mammalian expression plasmids of the extracellular domain of TLR2 and its two fragments were constructed. METHODS The cDNA fragment encoding complete extracellular domain was isolated by semi nested RT PCR methodwith the total RNA extracted from HL 60 cells, linked into pGEM T Easy vector, transformed into E.coli JM109, and inserted into pcDNA3 vector. Then two fragments amplified from it were linked into pGEM T Easy vector and pcDNA3. RESULTS The result of sequencing indicated that the sequences of our cDNA was identical to the corresponding parts of the mRNA for human TLR2 in GenBank except 4 nucleotides, and with a homology of 0.998. CONCLUSION The extracellular domain of TLR2 and its two fragments are cloned successfully.
作者 张林 于文彬 马越云 苏明权 丁振若
机构地区 第四军医大学西京医院检验科临床分子生物学研究室
出处 《第四军医大学学报》 北大核心 2002年第12期1085-1089,共5页 Journal of the Fourth Military Medical University
关键词 TOLL样受体 亮氨酸 重复序列 核酸 逆转录聚合酶链反应 克隆 分子 序列分析 Toll like receptor leucine repetitive sequences, nucleic acid reverse transcriptase polymerase chain reaction cloning, molecular sequence analysis
分类号 Q78 [生物学—分子生物学]
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共引文献3

同被引文献42

  • 1张炜,袁金铎,安利国.由不同接头分子介导的Toll样受体信号通路[J].细胞与分子免疫学杂志,2006,22(4):544-546. 被引量：5
  • 2罗冰,刘艳君,杨翠兰,富宁.TLR2在小鼠肿瘤细胞表面的表达[J].中国实验诊断学,2006,10(11):1245-1247. 被引量：2
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第四军医大学学报
2002年 第12期

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