Mouse high mobility group protein 1cDNA Cloning expression and identification of.doc

 PAGE \* MERGEFORMAT 10 Mouse high mobility group protein 1cDNA Cloning expression and identification of Of: Shi Yan Dai Xiaoli Yuan Xue Raymond Elite Wang Shengjun Suzhao Liang Chen Jianguo Xu summer of River St. Shaoqi Xiang Huang Xinxiang [Abstract] Objective: Cloning and prokaryotic expression of murine high-mobility group protein 1 (high mobility group box1, HMGB1. METHODS: The spleen cells extracted from mouse total RNA, RT PCR applied to obtain mice HMGB1cDNA, mice were amplified by PCR HMGB1 gene, the gene was constructed by TA cloning into the PMD 18T vector and sequencing, the gene was inserted into pET 28a (+) expression vector, IPTG induced, can be expressed relative molecular mass of approximately 30 kDa protein. blotting protein expression target protein. Results: RT PCR amplified 648 bp of DNA fragments, sequence analysis revealed that the known gene sequence database reported fully consistent with HMGB1 coding sequence was constructed prokaryotic expression vector, induced expression, Western blot identification, access to the molecular weight of about 30 kDa fusion protein. Conclusion: Cloning and expression of mouse HMGB1 genes, HMGB1 protein function tests for the foundation. [Key Words] high mobility group protein, cloning, prokaryotic expression [Abstract] Objective: To clone and express mouse high mobility group box chromosomal protein1 (HMGB1) gene.Methods: Total RNA was extracted from mouse spleen cells, and the whole length of HMGB1 gene was obtained by RT PCR.The HMGB1 gene was cloned into PMD 18T vector by TA clone and sequenced. Then the gene was inserted into pET 28a (+) expression vector. identified by Western Blot. Results: The target DNA sequence of HMGB1 was obtained by RT PCR which was about 648 bp length, sequencing results showed that HMGB1 gene was exactly consistent with the sequence reported in GenBank; Western Blot identified there was a protein strap about Mr.30 000.The HMGB1 protein was successfully expre