猪SEPW1基因的转录调控分析

doi:10.16431/j.cnki.1671-7236.2019.06.020

Abstract

Abstract:

This study was aimed to clone the potential promoter region of porcine SEPW1 gene and analyze its transcriptional activity,obtain its core promoter region,and analyze the effect of transcription factor SP1 on the transcriptional activity of SEPW1 gene for further studying the function of porcine SEPW1 gene in meat traits.The expression level of SEPW1 gene in Large White pigs tissues was detected by Real-time quantitative PCR,and the spatial expression profile was constructed.6 SEPW1 gene promoter deletion fragments were cloned by PCR,and 6 dual luciferase reporter vectors were constructed,the core promoter region of SEPW1 gene was obtained by detecting the dual luciferase activity of each vector;The potential transcription factor SP1 binding site was predicted through bioinformatics analysis of the core promoter region.The transcription factor SP1 binding site and its effect on the transcriptional activity of SEPW1 gene were confirmed by over-expression,down-expression,site-directed mutation and EMSA.The results showed that SEPW1 gene was widely expressed in 12 tissues of 4-month-old Large White pigs,while the expression levels were higher in castrocnemius and heart.The results of dual luciferase activity indicated that there was the core promoter region of SEPW1 gene 5'-flanking region at -443 to -231 bp,and there was a potential SP1 binding site at -378 to -306 bp.The results of over-expression and down-expression of SP1 gene indicated that transcription factor SP1 promoted the transcription of SEPW1 gene.Site-directed mutagenesis and EMSA assay confirmed that transcription factor SP1 could directly bind to SP1 binding sites (-348 to -339 bp) in the promoter region of SEPW1 gene.The results suggested that transcription factor SP1 could directly target the promoter region of SEPW1 gene and promote SEPW1 gene transcription.

Key words: porcine; SEPW1 gene; SP1; promoter; transcriptional activity

CLC Number:

S813.3

ZHANG Feng, LI Xin, CHEN Mingxin. Transcriptional Regulation Analysis of Porcine SEPW1 Gene[J]. , 2019, 46(6): 1730-1738.

References

[1] RAYMAN M P.Selenium in cancer prevention:A review of the evidence and mechanism of action[J].Proceedings of the Nutrition Society,2005,64(4):527-542.
[2] ZHU Y G, ELIZABETH A H,ZHAO F J,et al.Selenium in higher plants:Understanding mechanisms for biofortification and phytoremediation[J]. Trends in Plant Science,2009,14(8):436-442.
[3] FAIRWEATHER-TAIT S J,BAO Y,BROADLEY M R,et al.Selenium in human health and disease[J].Antioxidants & Redox Signaling,2011,14(7):1337-1383.
[4] 卢建新,张仲欣,任丽影.富硒食品加工现状与发展分析[J].农产品加工,2014,21:45-51. LU J X,ZHANG Z X,REN L Y.Processing status and development of selenium-accumlating food[J]. Academic Periodica of Farm Products Processing, 2014,21:45-51.(in Chinese)
[5] 王晓龙,许凯,秦藕菊,等.SelW基因转录后沉默对小鼠骨骼肌细胞内GSH及GPx影响研究[J].畜牧兽医学报,2009,40(7):1013-1018. WANG X L,XU K,QIN O J,et al.The influence of SelW on GSH and GPx after post-transcriptional gene silencing in mouse skeletal muscle cell[J].Acta Veterinaria et Zootechnica Sinica,2009,40(7):1013-1018.(in Chinese)
[6] 杨旭,董文宾.富硒食品的研究进展[J].食品安全质量检测学报,2017,8(6):2091-2097. YANG X,DONG W B.Research progress of Se-enriched food[J].Journal of Food Safety & Quality,2017,8(6):2091-2097.(in Chinese)
[7] 胡万明.富硒食品的研究意义与进展[J].食品安全导刊,2018,12:42. HU W M.Research significance and progress of selenium enriched food[J]. China Food Safety Magazine,2018,12:42.(in Chinese)
[8] JEONG D,KIM T,CHUNG Y,et al.Selenoprotein W is a glutathione-dependent antioxidant in vivo[J].FEBS Letter,2002,517(1-3):225-228.
[9] GU Q P,SUN Y,REAM L W,et al.Selenoprotein W accumulates primarily in primate skeletal muscle,heart,brain and tongue[J].Molecular and Cellular Biochemistry,2000,204(1-2):49-56.
[10] PRINZ W A,ASLUND F,HOLMGREN A,et al.The role of the thioredoxin and glutaredoxin pathways in reducing protein disulfide bonds in the Escherichia coli cytoplasm[J].The Journal of Biological Chemistry,1997,272(25):15661-15667.
[11] BELLINGHAM J,GREGORY-EVANS K,FOX M F,et al.Gene structure and tissue expression of human selenoprotein W,SEPW1,and identification of a retroprocessed pseudogene,SEPW1P[J].Biochimica et Biophysica Acta,2003,1627(2-3):140-146.
[12] LI J G,ZHOU J C,ZHAO H,et al.Enhanced water-holding capacity of meat was associated with increased Sepw1 gene expression in pigs fed selenium-enriched yeast[J].Meat Science,2011,87(2):95-100.
[13] LOFLIN J,LOPEZ N,WHANGER P D,et al.Selenoprotein W during development and oxidative stress[J].Journal of Inorganic Biochemistry,2006,100(10):1679-1684.
[14] DENG B,ZHANG F,CHEN K,et al.MyoD promotes porcine PPARgamma gene expression through an E-box and a MyoD-binding site in the PPARgamma promoter region[J].Cell and Tissue Research,2016,365(2):381-391.
[15] YEH J Y,BEILSTEIN M A,ANDREWS J S,et al.Tissue distribution and influence of selenium status on levels of selenoprotein W[J]. FASEB Journal,1995,9(5):392-396.
[16] GU Q,SUN Y,REAM L,et al.Selenoprotein W accumulates primarily in primate skeletal muscle,heart,brain and tongue[J].Molecular and Cellular Biochemistry,2000,204(1-2):49-56.
[17] BRAHMACHARY M,SCHONBACH C,YANG L,et al.Computational promoter analysis of mouse,rat and human antimicrobial peptide-coding genes[J].BMC Bioinformatics,2006,7(Suppl5):S8.
[18] LEE B J,PARK S I,PARK J M,et al.Molecular biology of selenium and its role in human health[J].Molecules and Cells,1996,6(5):509-520.
[19] REDERSTORFF M,KROL A,LESCURE A.Understanding the importance of selenium and selenoproteins in muscle function[J].Cellular and Molecular Life Sciences,2006,63(1):52-59.
[20] VENDELAND S C,BEILSTEIN M A,CHEN C L,et al.Purification and properties of selenoprotein W from rat muscle[J].The Journal of Biological Chemistry, 1993,268(23):17103-17107.
[21] 王晓龙,宋晓东,宋月峰,等.SelW mRNA下调对肌细胞状态影响的FCM评估[J].吉林农业大学学报,2007,29(2):152-154. WANG X L,SONG X D,SONG Y F,et al.Assessment of cell conditions by FCM after SelW mRNA down-regulation[J].Journal of Jilin Agricultural University,2007,29(2):152-154.(in Chinese)
[22] PEDERSEN N D,WHANGER P D,WESWIG P H,et al.Selenium binding proteins in tissues of normal and selenium responsive myopathic lambs[J].Bioinorganic Chemistry, 1973,2(1):33-45.

Transcriptional Regulation Analysis of Porcine SEPW1 Gene

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	CHEN Kaiwen, ZHAO Yunjiao, WANG Lei, ZHANG Jianbo, MA Yuhong, WU Guofang. Transcriptome Analysis of Porcine Leydig Cells Under Different Oxygen Concentrations [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2185-2195.
[2]	CAO Qi, JIN Qing, YUAN Yi, FENG Yixue, KONG Danni, CHEN Xianghong, WU Bin, SONG Wenbo, TANG Xibiao. Epidemiological Investigation of Common Respiratory Bacterial Pathogens in Large-scale Pig Farms [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2439-2449.
[3]	SHI Qiuying, DUAN Baomin, LI Ying, NIU Dengyun, GENG Chenyang, ZHOU Jiawei, WANG Rui, FENG Jingjing. Analysis of Genetic Variation of S1 Gene of Porcine Epidemic Diarrhea Virus in Some Areas of China from 2020 to 2021 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2468-2478.
[4]	OU Yunwen, PAN Qin, WANG Yang, DAI Junfei, REN Shaoke, ZHANG Yang, ZHANG Jie. Truncated Expression of ORF2 Gene of Porcine Parvovirus Type 6 and Preparation of Its Polyclonal Antibody [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2487-2495.
[5]	LI Mingguo, LI Qingchun, CHEN Xin, LU Shihao, HE Fan, QI Mengfan, ZHANG Huapeng, REN Yujun, ZHANG Qingze, FU Binbin, XU Mengsi, AI Zikai, YAN Kun, FENG Yun, HUA Zaidong, HUANG Tao, BI Yanzhen. Effects of TET Demethylase on Porcine Oocyte Development [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1907-1917.
[6]	KANG Pu, REN Zhaowen, CAI Rujian, LYU Zongji, HUANG Yuhao, WANG Xiaohu. Variation Regularity and Transmission of PRRSV-2 Lineage 8 Strains in Guangdong and Surrounding Provinces [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2001-2011.
[7]	JIANG Haijun, WANG Dan, GUO Yu, WEI Li, HOU Lei, WANG Jing, ZHU Shanshan, LIU Jue. Fusion Expression and Immunogenicity of Recombinant Cap-flagellin Protein of Porcine Circovirus Type 3 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2044-2052.
[8]	CHEN Tianbao, WANG Yani, LIU Baoling, WANG Gang, WANG Xiaohu, LIU Xuhui, CAI Rujian, HE Dongsheng. Research Progress on the Anti-Porcine Reproductive and Respiratory Syndrome Virus Effect and Mechanism of Traditional Chinese Medicine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2114-2129.
[9]	HUO Mingkai, GUAN Feihu, QIU Runhui, WEI Chunyan, YU Haitao, ZHU Jiale, ZHANG Hui. Study on the Mechanism of DDX1 Inhibiting Porcine Epidemic Diarrhea Virus Replication [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1556-1566.
[10]	LIU Jiaxing, HAN Xueying, ZHANG Xinru, DENG Jiaxin, YAO Lunguang, LIU Yangkun. Preparation and Immunogenicity Analysis of Ferritin Nanoparticles Carrying Antigenic Epitopes of Porcine Epidemic Diarrhea Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1567-1574.
[11]	ZHANG Guoqing, JIANG Yuhang, ZHANG Shuang, LI Letian, HAO Jiayi, CHEN Jing, ZOU Wancheng, LI Chang, LI Taiyuan, GAO Xu, JIN Ningyi. Preparation and Identification of PDCoV RBD Protein Peptide Antibody [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1575-1583.
[12]	SONG Xingchao, MENG Jinzhu, ZHAO Yuanyuan, WU Zhenyang, AN Qingming. Cloning and Bioinformatics Analysis of MyoG Gene Promoter Region Sequence in Guizhou White Goat [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 893-903.
[13]	LIU Xinyue, LI Lifang, WANG Zhenbao, ZHANG Wenliang, ZHANG Xin, ZHAO Hongtao, LI Pengyu, ZHENG Haotian, XIAO Jin, QI Peng. Adjuvants Screening and Immune Protection of Porcine Circovirus Type 2 Synthetic Peptide Vaccine [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1068-1080.
[14]	ZHANG Jingxu, XU Yu, LIANG Haiying, ZENG Zhiyong, TANG Deyuan, WANG Bin, XU Songping, WAN Juan, ZHU Yang. Phylogenetic Evolution and Epitope Analysis of Pseudorabies Virus Guizhou Isolates and Vaccine Strains [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1093-1106.
[15]	LI Huawei, GUO Kewei, WANG Xuying, ZHANG Xiaoyu, LU Ziwei, LI Xinfeng, MA Hui, HOU Wenjing. Eukaryotic Expression and Subcellular Localization of Nonstructural Protein 7 of Porcine Reproductive and Respiratory Syndrome Virus [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1160-1168.