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Expression, Purification and Bioactivities Analysis of Recombinant Active Peptide from Shark Liver
School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China
College of Life Sciences, Zhejiang Sci-Tech University, Hangzhou 310018, China
* Author to whom correspondence should be addressed.
Received: 20 April 2009; in revised form: 13 June 2009 / Accepted: 13 June 2009 / Published: 22 June 2009
Abstract: The Active Peptide from Shark Liver (APSL) was expressed in E. coli BL21 cells. The cDNA encoding APSL protein was obtained from shark regenerated hepatic tissue by RT-PCR, then it was cloned in the pET-28a expression vector. The expressed fusion protein was purified by Ni-IDA affinity chromatography. SDS-PAGE and HPLC analysis showed the purity of the purified fusion protein was more than 98%. The recombinant APSL (rAPSL) was tested for its biological activity both in vitro, by its ability to improve the proliferation of SMMC7721 cells, and in vivo, by its significant protective effects against acute hepatic injury induced by CCl4 and AAP (acetaminophen) in mice. In addition, the rAPSL could decrease the blood glucose concentration of mice with diabetes mellitus induced by alloxan. Paraffin sections of mouse pancreas tissues showed that rAPSL (3 mg/kg) could effectively protect mouse islets from lesions induced by alloxan, which indicated its potential application in theoretical research and industry.
Keywords: APSL; prokaryotic expression; bioactivity analysis
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MDPI and ACS Style
Lv, Z.; Ou, Y.; Li, Q.; Zhang, W.; Ye, B.; Wu, W. Expression, Purification and Bioactivities Analysis of Recombinant Active Peptide from Shark Liver. Mar. Drugs 2009, 7, 258-267.
Lv Z, Ou Y, Li Q, Zhang W, Ye B, Wu W. Expression, Purification and Bioactivities Analysis of Recombinant Active Peptide from Shark Liver. Marine Drugs. 2009; 7(2):258-267.
Lv, Zhengbing; Ou, Yu; Li, Qian; Zhang, Wenping; Ye, Boping; Wu, Wutong. 2009. "Expression, Purification and Bioactivities Analysis of Recombinant Active Peptide from Shark Liver." Mar. Drugs 7, no. 2: 258-267.