水牛STAT4基因克隆分析及其真核表达载体的构建

doi:10.16431/j.cnki.1671-7236.2016.08.001

Abstract

Abstract:

In order to elucidate the function and molecular mechanisms of buffalo signal transducer and activator of transcription 4 (STAT4) gene during folliculogenesis,embryogenesis and lactogenesis,buffalo STAT4 gene was studied with 3' -RACE,bio-informatics analysis,eukaryotic vector construction and cell transfection technologyin this study.The results showed that the coding region of buffalo STAT4 was 2 247 bp,3'-was 268 bp,and encoded 748 amino acids;BLAST analysis showed that the buffalo STAT4 gene shared 99%,99%,99%,95%,93%,93% and 92% of similar nucleotide sequence with that of Bos taurus, Capra hircus,Ovis aries,Sus scrofa,Equus caballus,Canis lupus and Homo sapiens,respectively.STAT4 protein was weakly acidic,without signal peptide,located in the cytoplasmic,and with the presence of STAT_int,STAT_alpha,STAT_bind and SH2_STAT4 domain.bta-miR-200a,bta-miR-2429 and bta-miR-2410 and so on were potential microRNAs of STAT4 3'-UTR.The buffalo STAT4 eukaryotic expression vector was successfully constructed,after transfected into HEK293T cell lines,STAT4-EGFP fusion protein was detectable.

Key words: buffalo; STAT4 gene; cloning; bio-informatics analysis; vector construction

CLC Number:

ZHU Peng, PANG Chun-ying, DUAN An-qin, DENG Ting-xian, LU Xing-rong, LIANG Xian-wei. Cloning and Construction of Eukaryotic Expression Vector of Buffalo STAT4 Gene[J]. , 2016, 43(8): 1929-1937.

References

[1] Wang Y,Qu A,Wang H.Signal transducer and activator of transcription 4 in liver diseases[J].Int J Biol Sci,2015,11(4):448-455.
[2] Frucht D M,Aringer M,Galon J,et al.STAT4 is expressed in activated peripheral blood monocytes,dendritic cells,and macrophages at sites of Th1-mediated inflammation[J]. J Immunol,2000,164(9):4659-4664.
[3] Yamamoto K,Kobayashi H,Arai A,et al.cDNA cloning,expression and chromosome mapping of the human STAT4 gene:Both STAT4 and STAT1 genes are mapped to 2q32.2-->q32.3[J].Cytogenet Cell Genet,1997,77(3-4):207-210.
[4] Barnes J,Agarwal S K.Targeting STAT4 in systemic sclerosis:A promising new direction[J].Expert Rev Clin Immunol,2011,7(4):445-448.
[5] Wang C,Sandling J K,Hagberg N,et al.Genome-wide profiling of target genes for the systemic lupus erythematosus-associated transcription factors IRF5 and STAT4[J].Ann Rheum Dis,2013,72(1):96-103.
[6] Choi K S,Scorpio D G,Dumler J S.STAT1 negatively regulates immune-mediated injury with Anaplasma phagocytophilum infection[J].J Immunol, 2014,193(10):5088-5098.
[7] Enomoto D,Obana M,Miyawaki A, et al.Cardiac specific ablation of STAT3 gene in subacute phase of myocardial infarction exacerbated cardiac remodeling[J].Am J Physiol Heart Circ Physiol,2015,309(3):H471-H480.
[8] Takeda K,Noguchi K,Shi W, et al.Targeted disruption of the mouse STAT3 gene leads to early embryonic lethality[J].Proc Natl Acad Sci USA,1997,94(8):3801-3804.
[9] McWilliams I L,Rajbhandari R,Nozell S,et al.STAT4 controls GM-CSF production by both Th1 and Th17 cells during EAE[J].J Neuroinflammation,2015,12:128.
[10] Shi A,Dong J,Hilsenbeck S,et al.The status of STAT3 and STAT5 in human breast atypical ductal hyperplasia[J].PLoS One,2015,10(7):e0132214.
[11] Liu X,Robinson G W,Wagner K U, et al.STAT5A is mandatory for adult mammary gland development and lactogenesis[J].Genes Dev,1997,11(2):179-186.
[12] Edukulla R,Singh B,Jegga A G,et al.Th2 cytokines augment IL-31/IL-31RA interactions via STAT6-dependent IL-31RA expression[J].J Biol Chem,2015,290(21):13510-13520.
[13] Haricharan S,Li Y.STAT signaling in mammary gland differentiation,cell survival and tumorigenesis[J].Mol Cell Endocrinol,2014,382(1):560-569.
[14] Philp J A,Burdon T G,Watson C J.Differential activation of STATs 3 and 5 during mammary gland development[J].FEBS Lett,1996,396(1):77-80.
[15] 朱鹏,粟小平,苏节,等.沼泽型水牛ALK3基因克隆分析与表达模式研究[J].中国畜牧兽医,2015,42(3):516-524.
[16] 苏节,朱鹏,刘庆友,等.水牛转录抑制因子CTCF基因克隆分析及不同组织中的表达研究[J].中国畜牧兽医,2013,40(3):1-7.
[17] 苏节,刘庆友,朱鹏,等.沼泽型水牛CYP19A1基因克隆、序列分析及组织表达研究[J].畜牧兽医学报,2013,44(4):514-521.
[18] Letunic I,Doerks T,Bork P.SMART 7:Recent updates to the protein domain annotation resource[J].Nucleic Acids Res,2012,40(Database issue):D302-D305.
[19] Petersen T N,Brunak S,von Heijne G,et al.Signal P 4.0:Discriminating signal peptides from transmembrane regions[J].Nat Methods,2011,8(10):785-786.
[20] 邓廷贤,庞春英,王建,等.水牛分子生物学技术研究新进展[J].中国畜牧兽医,2014,41(4):79-84.
[21] Huang Y,Lei Y,Zhang H,et al.microRNA regulation of STAT4 protein expression:Rapid and sensitive modulation of IL-12 signaling in human natural killer cells[J].Blood,2011,118(26):6793-6802.
[22] Song X M,Zhang L,Jiang J F,et al.An Sdu Ⅰ polymorphism at intron 20 of the Chinese Holstein cow STAT4 gene and its effect on milk performance traits[J].Genet Mol Res,2013,12(2):1593-1602.
[23] Zhang F,Huang J,Li Q,et al.Novel single nucleotide polymorphisms (SNPs) of the bovine STAT4 gene and their associations with production traits in Chinese Holstein cattle[J].AJB,2010,9:4003-4008.

Cloning and Construction of Eukaryotic Expression Vector of Buffalo STAT4 Gene

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	XU Yuanyuan, ZHENG Haiying, YANG Chunyan, DENG Tingxian, HUANG Chenqian, FENG Chao, LU Xingrong, DUAN Anqin, MO Xia, MA Xiaoya, SHANG Jianghua. Effects of Inhibition of BMP/Smad Signaling Pathway on the Growth and Steroid Hormone Synthesis of Buffalo Ovarian Granulosa Cells [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2354-2362.
[2]	XIAO Peng, SHANG Jianghua, YANG Chunyan, LI Mengqi, DUAN Anqin, MA Xiaoya, FENG Chao, HUANG Chenqian, ZHANG Bo, ZHOU Jinchen, WEI Kelong, ZHENG Wei, ZHENG Haiying. Effects of Alpha-linolenic Acid on Buffalo Ovarian Granulosa Cells Cultured in vitro [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2380-2387.
[3]	FAN Bingfeng, LI Wen, LIU Lixiang, ZHAO Xiangyuan, SHAO Jing, LIN Lefeng, SUN Huimin, ZHANG Xulin, XU Baozeng. Cloning and Bioinformatics Analysis of AANAT Gene in Mink [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1307-1318.
[4]	LIU Hongrun, ZHU Siran, FENG Lingli, ZHANG Kun, YAN Gang, WANG Yubin, ZHANG Shuai, JIANG Shan, XU Di, LAN Ganqiu, LIANG Jing. Cloning,Bioinformatics Analysis and Tissue Expression Localization of CDO1 Gene in Large White Pigs [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1352-1363.
[5]	ZHANG Yongjie, SUI Zhiyuan, ZHANG Zhishuai, WANG Chenguang, LI Xiaojun, XING Feng. Cloning and Bioinformatics Analysis of NPTX1 Gene and Its Tissues Expression at Puberty in Dolang Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1048-1058.
[6]	WEI Yirong, ZHENG Zihua, ZHANG Sanbao, SONG Ying, JIANG Haiyu, SUN Wenyue, CHENG Pengfei, LIU Yufan, ZOU Jianwei, HUANG Yanna, PAN Yan, JIANG Qinyang. Cloning,Bioinformatics Analysis and Tissue Expression of UCP1 Gene in Nubian Goats [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(2): 440-450.
[7]	TANG Jingyu, DU Hanyu, MENG Chunchun, LIU Guangqing. Cloning,Bioinformatics Analysis and Subcellular Localization of Interferon-stimulated Gene 15 in Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(8): 2843-2854.
[8]	QI Nannan, QIAN Yonghang, LUO Gang, SITU Xuqi, LIU Jiying. Cloning, Bioinformatics Analysis and Tissue Expression Location of Porcine MORC2 Gene [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2431-2441.
[9]	ZHANG Hang, SHA Huiyang, LI Huawei, HUANG Liangzong, ZHAO Mengmeng. Cloning, Bioinformatics and Tissue Expression Analysis of TRIM3 Gene in Swine [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(7): 2474-2483.
[10]	YAN Yelian, ZHANG Mengya, LIU Qiuchen, WANG Xin, XU Changzhi, ZONG Yanfeng, ZHU Zhihua, WU Sucheng, SONG Yu, LI Yunsheng, ZHANG Yunhai, CAO Zubing. Research Progress of Epigenetic Reprogramming in Mammalian Somatic Cell Nuclear Transfer Embryo Development [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(6): 2259-2269.
[11]	WANG Baobao, JIN Hang, XIAN Yuhan, FENG Hongsheng, GAO Fengshan. Cloning SLA-1 Gene from Porcine Kidney Epithelial Cells and Analyzing its Molecular Structure Characteristics [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1671-1678.
[12]	ZHANG Jihu, MI Renisa, WANG Zixuan, LI Lin, XING Feng, LIU Shudong. Cloning and Bioinformatics of ERα Gene and Its Expression of Ovary Tissue in Cele Black Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(5): 1679-1687.
[13]	SHEN Xiang, CHEN Linghui, YAO Jingjie, WANG Sanhu, BAI Yueyu, ZHANG Xiaojian. Cloning,Bioinformatics and Expression Analysis of Bovine CCR7 Gene [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1244-1252.
[14]	ZHANG Zhishuai, SUI Zhiyuan, ZHANG Jihu, LI Qingjin, XING Feng. Cloning of CNP Gene and Its Tissue Expression Characteristics at Puberty in Dolang Sheep [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(4): 1402-1412.
[15]	HOU Jinxing, WANG Zhanhang, ZHU Junru, JIANG Yue, LIU Shujuan, AN Xiaopeng. Cloning, Bioinformatics and Tissue Expression Profile Analysis of CSN1S1 Gene in Guanzhong Dairy Goats [J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(3): 857-865.