猪lipasin基因的克隆及其对肝细胞LPL基因转录的影响

doi:10.11843/j.issn.0366-6964.2016.02.027

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (2): 411-416.doi: 10.11843/j.issn.0366-6964.2016.02.027

• 研究简报 • 上一篇

猪lipasin基因的克隆及其对肝细胞LPL基因转录的影响

李曼曼，温丙言，朱河水，钟凯，杨国宇，王月影^*

（河南农业大学农业部动物生化与营养重点开放实验室，郑州 450002）

收稿日期:2015-03-18 出版日期:2016-02-23 发布日期:2016-02-23
通讯作者: 王月影，副教授，硕士生导师，E-mail：wangyueying2008@126.com
作者简介:李曼曼（1989-），女，河南许昌人，硕士生，主要从事动物生物技术方面的研究，E-mail：1065857330@qq.com
基金资助:
农业部“948”重点计划（2011-G35）；农业部转基因重大专项（2014ZX0801015B）；河南省重点科技攻关项目（112102310705）

Cloning of Porcine lipasin Gene and Its Effect on LPL Gene Transcription in Liver Cell

LI Man-man，WEN Bing-yan，ZHU He-shui，ZHONG Kai，YANG Guo-yu，WANG Yue-ying^*

(Key Laboratory of Animal Biochemistry and Nutrition of Ministry of Agriculture，Henan Agricultural University，Zhengzhou 450002，China)

Received:2015-03-18 Online:2016-02-23 Published:2016-02-23

摘要/Abstract

摘要：

旨在克隆猪lipasin 基因，并检测其对肝细胞LPL基因转录的影响。本试验利用同源电子克隆技术克隆猪lipasin基因，构建其真核表达载体转入肝细胞，探讨其对LPL基因转录的影响。结果表明，成功克隆了猪lipasin基因，目的片段大小为615 bp，上传至GenBank(登录号：KF017598)，开放阅读框为594 bp，编码197个氨基酸，liapsin蛋白的分子量和等电点分别为21.99 ku和6.79；成功构建了真核表达载体PB-EGFP-lipasin，实现lipasin基因在肝细胞中的过表达，与对照组相比，PB-EGFP-lipasin转染的细胞中LPL基因的转录水平显著下降。结果提示，猪lipasin可能通过抑制肝细胞LPL的转录影响脂代谢。

Abstract:

The study aimed to clone the lipasin gene and investigate its effects on LPL gene transcription in liver cell in pig.The porcine lipasin gene was obtained by using homologous electronic cloning techniques and the eukaryotic expression vector was constructed and transfected into hepatocytes，and further to identify its effect on the transcription level of LPL gene.The results showed that lipasin gene was successfully cloned，the segment size was 615 bp，it was deposited to GenBank with the accession number KF017598.The size of open reading frame (ORF) of lipasin was 594 bp and it encoded 197 amino acids.The weight of lipasin molecular and theoretical isoelectric point were 21.99 ku and 6.79，respectively.The recombinant eukaryotic expression vector PB-EGFP-lipasin was successfully constructed and transfected into hepatocyte，realizing the overexpression of lipasin gene.Compared with the control group，the transcription level of LPL mRNA decreased significantly in PB-EGFP-lipasin transfected hepatocyte.These results suggest that lipasin may affect lipid metabolism by inhibiting the LPL gene transcription in hepatocyte.

中图分类号:

S828
S813.3

李曼曼，温丙言，朱河水，钟凯，杨国宇，王月影. 猪lipasin基因的克隆及其对肝细胞LPL基因转录的影响[J]. 畜牧兽医学报, 2016, 47(2): 411-416.

LI Man-man，WEN Bing-yan，ZHU He-shui，ZHONG Kai，YANG Guo-yu，WANG Yue-ying. Cloning of Porcine lipasin Gene and Its Effect on LPL Gene Transcription in Liver Cell[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(2): 411-416.

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参考文献

［1］MATTIJSSEN F，KERSTEN S.Regulation of triglyceride metabolism by Angiopoietin-like proteins［J］.Biochim Biophys Acta, 2012，1821(5)：782-789.
［2］ONO M，SHIMIZUGAWA T，SHIMAMURA M，et al.Protein region important for regulation of lipid metabolism in angiopoietin-like 3 (ANGPTL3)：ANGPTL3 is cleaved and activated in vivo［J］.J Biol Chem, 2003，278(43)：41804-41809.
［3］CAMENISCH G，PISABARRO M T，SHERMAN D，et al.ANGPTL3 stimulates endothelial cell adhesion and migration via integrin alpha vbeta 3 and induces blood vessel formation in vivo［J］. J Biol Chem, 2002，277(19)：17281-17290.
［4］KOISHI R，ANDO Y，ONO M，et al.Angptl3 regulates lipid metabolism in mice［J］. Nat Genet，2002，30(2)：151-157.
［5］LEE E C，DESAI U，GOLOLOBOV G，et al.Identification of a new functional domain in angiopoietin-like 3 (ANGPTL3) and angiopoietin-like 4 (ANGPTL4) involved in binding and inhibition of lipoprotein lipase (LPL)［J］.J Biol Chem, 2009，284(20)：13735-13745.
［6］KOSTER A，CHAO Y B，MOSIOR M，et al.Transgenic angiopoietin-like (angptl)4 overexpression and targeted disruption of angptl4 and angptl3：regulation of triglyceride metabolism［J］.Endocrinology, 2005，146(11)：4943-4950.
［7］MIIDA T，HIRAYAMA S.Impacts of angiopoietin-like proteins on lipoprotein metabolism and cardiovascular events［J］.Curr Opin Lipidol, 2010，21(1)：70-75.
［8］FU Z，YAO F，ABOU-SAMRA A B，et al.Lipasin，thermoregulated in brown fat，is a novel but atypical member of the angiopoietin-like protein family［J］.Biochem Biophys Res Commun, 2013，430(3)：1126-1131.
［9］ZHANG R.Lipasin，a novel nutritionally-regulated liver-enriched factor that regulates serum triglyceride levels［J］.Biochem Biophys Res Commun,2012，424(4)：786-792.
［10］REN G，KIM J Y，SMAS C M.Identification of RIFL，a novel adipocyte-enriched insulin target gene with a role in lipid metabolism［J］.Am J Physiol Endocrinol Metab, 2012，303(3)：E334-351.
［11］TANG T，LI L，TANG J，et al.A mouse knockout library for secreted and transmembrane proteins［J］.Nat Biotechnol，2010，28(7)：749-755.
［12］ZHANG R，ABOUSAMRA A B.Emerging roles of Lipasin as a critical lipid regulator［J］.Biochem Biophys Res Commun,2013，432(3)：401-405.
［13］QUAGLIARINI F，WANG Y，KOZLITINA J，et al.Atypical angiopoietin-like protein that regulates ANGPTL3［J］. Proc Natl Acad Sci, 2012，109(48)：19751-19756.
［14］PATSCH J R，MIESENBOCK G，HOPFERWIESER T，et al.Relation of triglyceride metabolism and coronary artery disease.Studies in the postprandial state［J］.Arterioscler Thromb, 1992，12(11)：1336-1345.
［15］KERSTEN S.Physiological regulation of lipoprotein lipase［J］.Biochim Biophys Acta, 2014，1841(7)：919-933.
［16］COLEMAN T，SEIP R L，GIMBLE J M，et al.COOH-terminal disruption of lipoprotein lipase in mice is lethal in homozygotes，but heterozygotes have elevated triglycerides and impaired enzyme activity［J］.J Biol Chem,1995，270(21)：12518-12525.
［17］ZHANG R，ABOU-SAMRA A B.A dual role of lipasin (betatrophin) in lipid metabolism and glucose homeostasis consensus and controversy［J］.Cardiovasc Diabetol, 2014，13(1)：133-141.

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猪lipasin基因的克隆及其对肝细胞LPL基因转录的影响

Cloning of Porcine lipasin Gene and Its Effect on LPL Gene Transcription in Liver Cell

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