基因表达_____高迁移率族蛋白1；试剂盒，诊断；基因表达

HMGB1 B盒编码序列的克隆和大肠杆菌表达

http://www.100md.com 《第四军医大学学报》 2005年第3期

基因表达,,高迁移率族蛋白1；试剂盒，诊断；基因表达,HMGB1B盒编码序列的克隆和大肠杆菌表达,0引言,1材料和方法,2结果,3讨论

     Cloning and expression of mouse HMGB1 B box coding sequence

    JIANG NanYan1, YU WenBin1, SU MingQuan1, LI Bin1, HAO XiaoKe1, LI LiWen2

    1Centre of Clinical Molecular Biology, 2Institute of Orthopaedics of Chinese PLA, Xijing Hospital, Fourth Military Medical University, Xian 710033, China

    【Absract】 AIM: To clone and express mouse High mobility group box 1 (HMGB1) B box coding sequence. METHODS: Total RNA was extracted from the lung tissue of a newborn mouse and the HMGB1 B box coding sequence was obtained by RTPCR using specific primers. The B box coding sequence was cloned into BamHI and EcoRI site of pGEX4T2 expression vector and confirmed by sequencing. After transforming E.coli BL21(DE3), the recombinant bacteria was induced at 30℃ for 5 h, the fusion protein GSTB box was analyzed by SDSPAGE. RESULTS: DNA sequencing results showed that the mouse HMGB1 B box coding sequence was exactly consistent with the sequence reported in GenBank, and the SDSPAGE analysis demonstrated that HMGB1 B box protein was expressed in E.coli BL21 (DE3). The protein band amounted to 30% of total bacteria protein. CONCLUSION: Mouse HMGB1 B box coding sequence is successfully cloned and expressed in E.coli. ......

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