title: Cloning and prokaryotic expression of the porcine lipasin gene.
abstract: Lipasin has recently been demonstrated to be involved in lipid metabolism. In this study, two specific primers were used to amplify the lipasin open reading frame from porcine liver tissue. The polymerase chain reaction product was cloned to a pGEM®-T Easy Vector, digested by SalI and NotI, and sequenced. The lipasin fragment was then cloned to a pET21(b) vector and digested by the same restriction enzyme. The recombinant plasmid was transferred to Escherichia coli (BL21), and the lipasin protein was induced with isopropyl-β-D-thiogalactopyranoside. The protein obtained was identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis and western blotting. A pET-lipasin prokaryotic recombinant expression vector was successfully constructed, and a 25.2-kDa protein was obtained. This study provides a basis for further research on the biological function of porcine lipasin.
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China.
authors.forename: M M
authors.initials: MM
authors.lastname: Li
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China.
authors.forename: J
authors.initials: J
authors.lastname: Geng
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China.
authors.forename: Y J
authors.initials: YJ
authors.lastname: Guo
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China.
authors.forename: X Q
authors.initials: XQ
authors.lastname: Jiao
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China.
authors.forename: W F
authors.initials: WF
authors.lastname: Lu
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China.
authors.forename: H S
authors.initials: HS
authors.lastname: Zhu
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China wangyueying2008@126.com.
authors.forename: Y Y
authors.initials: YY
authors.lastname: Wang
authors.affiliation: Key Laboratory of Animal Biochemistry and Nutrition, Ministry of Agriculture, Henan Agricultural University, Zhengzhou, Henan, China.
authors.forename: G Y
authors.initials: GY
authors.lastname: Yang
mesh_terms: D000091306:Angiopoietin-Like Protein 8; D000075802:Angiopoietin-like Proteins; D000818:Animals; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D050356:Lipid Metabolism / Q000235:genetics*; D008099:Liver / Q000378:metabolism; D036361:Peptide Hormones / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D013552:Swine / Q000235:genetics*
chemical_list: C576683:ANGPTL8 protein, human; D000091306:Angiopoietin-Like Protein 8; D000075802:Angiopoietin-like Proteins; D036361:Peptide Hormones; D011993:Recombinant Fusion Proteins