Cloning and Expression of SREBP-1 andSREBP-2 in Torafugu Takifugu rubripes

doi:10.7668/hbnxb.2016.04.013

Abstract

Abstract: In order to study the function and molecular mechanism of SREBP-1 and SREBP-2, the open reading frames (ORF) of torafugu SREBP-1 and SREBP-2 genes were amplified from total RNA of adipose tissue by reverse transcription-polymerase chain reaction (RT-PCR).The results showed that the ORF of SREBP-1 was composed of 3 330 bp,encoding 1 109 amino acids,while the ORF of SREBP-2 was composed of 3 444 bp,encoding 1 147 amino acids.The expression of SREBP-1 gene in torafugu tissues was determined using Real-time quantitative PCR,presenting that the highest expression was in brain,followed in eye,adipose tissue,spleen,gill,intestine,heart and liver,but the lowest in muscle.The prokaryotic expression system of recombined vector pET32a-SREBP-1 was constructed successfully.SDS-PAGE analysis showed that the expressed protein accumulated and the molecular weight of expressed fusion protein was 141 kDa.

Key words: Takifugu rubripes, SREBP-1 gene, SREBP-2 gene, Prokaryotic expression

CLC Number:

S917
Q786

ZHANG Wei, JI Mingli, GUO Qianjin, QIAN Zhibin. Cloning and Expression of SREBP-1 andSREBP-2 in Torafugu Takifugu rubripes[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2016, 31(4): 74-79. doi: 10.7668/hbnxb.2016.04.013.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.2016.04.013

http://www.hbnxb.net/EN/Y2016/V31/I4/74

References

[1] Wei S,Espenshade P.Expanding roles for SREBP in metabolism[J].Cell Metabolism,2012,16(4):414-419.
[2] Bennett M K,Ngo T T,Athanikar J N,et al.Co-stimulation of promoter for low density lipoprotein receptor gene by sterol regulatory element-binding protein and Sp1 is specifically disrupted by the yin yang 1 protein[J].The Journal of Biological Chemistry,1999,274(19):13025-13032.
[3] Deng X,Cagen L M,Wilcox H G,et al.Regulation of the rat SREBP-lc promoter in primary rat hepatocytes[J].Biochemical and Biophysical Research Communications,2002,290(1):256-262.
[4] Hua X,Yokoyama C,Wu J,et al.SREBP-2,a second basic-helix-loop-helix-leucine zipper protein that stimulates transcription by binding to a sterol regulatory element[J].Proceedings of the National Academy of Sciences of the United States of America,1993,90(24):11603-11607.
[5] Jeon T I,Osborne T F.SREBPs:metabolic integrators in physiology and metabolism[J].Trends in Endocrinology & Metabolism,2011,23(2):65-72.
[6] Brown M S,Goldstein J L.The SREBP pathway:regulation of cholesterol metabolism by proteolysis of a membrane-bound transcription factor[J].Cell,1997,89(3):331-340.
[7] Assaf S,Hazard D,Pitel F,et al.Cloning of cDNA encoding the nuclear form of chicken sterol response element binding protein-2 (SREBP-2),chromosomal localization,and tissue expression of chicken SREBP-1 and -2 genes[J].Poultry Science,2003,82(1):54-61.
[8] 张伟,付云,张煜.红鳍东方鲀SREBP-1 基因的电子克隆及生物信息学分析[J].广东农业科学,2014,41(17):119-123,241.
[9] Horton J D,Goldstein J L,Brown M S.SREBPs:activators of the complete program of cholesterol and fatty acid synthesis in the liver[J].The Journal of Clinical Investigation,2002,109(9):1125-1131.
[10] Tanaka N,Zhang X,Sugiyama E,et al.Eicosapentaenoic acid improves hepatic steatosis independent of PPARα activation through inhibition of SREBP-1 maturation in mice[J].Biochemical Pharmacology,2010,80(10):1601-1612.
[11] Ma P C,Rould M A,Weintraub H,et al.Crystal structure of MyoD bHLH domain-DNA complex:perspectives on DNA recognition and implications for transcriptional activation[J].Cell,1994,77(3):451-459.
[12] Shimomura I,Shimano H,Horton J D,et al.Differential expression of exons 1a and 1c in mRNAs for sterol regulatory element binding protein-1 in human and mouse organs and cultured cells[J].The Journal of Clinical Investigation,1997,99(5):838-845.
[13] Felder T K,Klein K,Patsch W,et al.A novel SREBP-1 splice variant:tissue abundance and transactivation potency[J].Biochimica et Biophysica acta,2005,1731(1):41-47.
[14] Harada N,Yonemoto H,Yoshida M,et al.Alternative splicing produces a constitutively active form of human SREBP-1[J].Biochemical and Biophysical Research Communications,2008,368(3):820-826.
[15] Minghetti M,Leaver,M J,Tocher D R.Transcriptional control mechanisms of genes of lipid and fatty acid metabolism in the Atlantic salmon (Salmo salar L.) established cell line,SHK-1[J].Biochim Biophys Acta,2011,1811(3):194-202.
[16] Dong X,Xu H,Mai K,et al.Cloning and characterization of SREBP-1 and PPAR-α in Japanese seabass Lateolabrax japonicus,and their gene expressions in response to different dietary fatty acid profiles[J].Comparative Biochemistry and Physiology,Part B,2015,180(180C):48-56.
[17] Zheng J L,Luo Z,Zhu Q L,et al.Molecular cloning and expression pattern of 11 genes involved in lipid metabolism in yellow catfish Pelteobagrus fulvidraco[J].Gene,2013,531(1):53-63.
[18] Pai W Y,Hu C C,Lai C Y,et al.Cannabinoid receptor 1 promotes hepatic lipid accumulation and lipotoxicity through the induction of SREBP-1c expression in zebrafish[J].Transgenic Research,2013,22(4):823-838.
[19] 刘芳.黄斑蓝子鱼固醇调节元件结合蛋白-1基因克隆及表达研究[D].汕头:汕头大学,2011.
[20] 王丽,那威,王宇祥,等.鸡SREBP1 基因抗血清制备及组织表达特性分析[J].细胞与分子免疫学杂志,2010,26(12):1241-1245.

Please choose a citation manager

Content to export