Expression, purification and bioactivities analysis of recombinant active peptide from shark liver.
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 CCl(4) 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.
Lv
Zhengbing
Z
School of Life Science and Technology, China Pharmaceutical University, Nanjing 210009, China.
Ou
Yu
Y
Li
Qian
Q
Zhang
Wenping
W
Ye
Boping
B
Wu
Wutong
W
D000818:Animals; D001786:Blood Glucose / Q000187:drug effects; D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D003001:Cloning, Molecular; D003921:Diabetes Mellitus, Experimental / Q000473:pathology; D005786:Gene Expression Regulation / Q000187:drug effects; D022781:Hepatocytes / Q000187:drug effects; D008099:Liver / Q000737:chemistry* / Q000187:drug effects; D051379:Mice; D010179:Pancreas / Q000187:drug effects; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000378:metabolism* / Q000494:pharmacology; D012754:Sharks / Q000502:physiology*
D001786:Blood Glucose; D011994:Recombinant Proteins