鸡Smad4基因克隆、生物信息学及组织表达分析

doi:10.16431/j.cnki.1671-7236.2021.10.003

Abstract

Abstract: The purpose of this study was to clone Smad4 gene and analyze its bioinformatics and tissue expression in chicken. Suqin No. 3 chicken was selected as experimental subjects, the full-length sequence of Smad4 gene in chicken was amplified by Race technology and cloned, the bioinformatics analysis of Smad4 gene was carried out, and the phylogenetic tree was constructed. The expression of Smad4 gene in chicken tissues was detected by Real-time quantitative PCR. The results showed that a full-length of Smad4 gene in chicken was obtained, which contained 17 bp 5'UTR, 1 842 bp open reading frame and 466 bp 3'UTR. There were 11 exons and 10 introns, the length of mRNA was 2 325 bp, which encoded 613 amino acids. The phylogenetic tree showed that Smad4 gene in chicken was clustered with other birds. Bioinformatics analysis showed that the molecular mass of Smad4 protein was 65.43 ku, the theoretical isoelectric point was 10.04, it was a hydrophilic protein. Smad4 protein contained two conserved domains MH1 and MH2. The secondary structure was consist of alpha helix (18.11%), extended chain (17.29%) and random coil (64.60%). Tissue expression analysis showed that Smad4 gene was highly expressed in heart, liver, jejunum and ovary, but there was no expression in breast muscle. In this study, the full-length sequence of Smad4 gene in chicken was successfully obtained, and its tissue expression pattern was preliminarily studied, which provided a theoretical basis for further study on the molecular mechanism of Smad4 gene in reproductive activity of chicken.

Key words: chicken; Smad4 gene; cloning; expression analysis

CLC Number:

HUANG Zhengyang, WANG Qianbao, LI Chunmiao, HUANG Huayun, LIANG Zhong, MU Chunyu, LI Shoufeng, ZHAO Zhenhua. Cloning, Bioinformatics and Tissue Expression Analysis of Smad4 Gene in Chicken (Gallus gallus)[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(10): 3522-3532.

References

[1] ZHAO M, MISHRAL, DENG C X.The role of TGF-β/SMAD4 signaling in cancer[J]. International Journal of Biological Sciences, 2018, 14(2):111-123.
[2] LI Z, ZHU M X, HU B, et al.Effects of suppressing Smads expression on wound healing in Hyriopsis cumingii[J]. Fish & Shellfish Immunology, 2020, 97:455-464.
[3] ZHU G, FANG C, LI J, et al.Transcriptomic diversification of granulosa cells during follicular development in chicken[J]. Scientific Reports, 2019, 9(1):5462.
[4] SILVESTRI L, NAI A, DULJA A, et al.Hepcidin and the BMP-SMAD pathway:An unexpected liaison[J]. Vitamins and Hormones, 2019, 110:71-99.
[5] LI X, TRIPURANI S K, JAMES R, et al.Minimal fertility defects in mice deficient in oocyte-expressed Smad4[J]. Biology of Reproduction, 2012, 86(1):1-6.
[6] LIM J, TU X, CHOI K, et al.BMP-Smad4 signaling is required for precartilaginous mesenchymal condensation independent of Sox9 in the mouse[J]. Developmental Psychobiology, 2015, 400(1):132-138.
[7] NI J, WU Q Q, LIAO H H, et al.Bcl6 suppresses cardiac fibroblast activation and function via directly binding to Smad4[J]. Current Medical Science, 2019, 39(4):534-540.
[8] 采复拉·大木拉, 田志龙, 段新华, 等.BMP4和SMAD4基因在公绵羊繁殖相关组织的表达[J]. 中国草食动物科学, 2019, 39(2):5-8. CAIFULA D, TIAN Z L, DUAN X H, et al.Expression of BMP4 and SMAD4 genes in ram reproduction-related tissues[J]. China Herbivores Science, 2019, 39(2):5-8.(in Chinese)
[9] 冯玮.骨细胞Smad4通过Notch信号调控成骨分化的机制研究[D].重庆:重庆医科大学, 2020. FENG W.Mechanisms of osteocytic Dmad4 regulating osteogenic differentiation through notch signaling[D].Chongqing:Chongqing Medical University, 2020.(in Chinese)
[10] 盖玉强, 杜宝霞, 马龙, 等.不同发情时期Smad4基因在绵羊卵巢中表达规律的研究[J]. 吉林畜牧兽医, 2020, 41(10):1-2. GAI Y Q, DU B X, MA L, et al.Expression study of Smad4 gene in ovine ovary at different estrus stages[J]. Jilin Animal Husbandry and Veterinary Medicine, 2020, 41(10):1-2.(in Chinese)
[11] 王玉恒, 徐业芬, 牛家强, 等.牦牛Smad4基因mRNA组织表达谱及其miRNAs初步研究[J]. 畜牧兽医学报, 2017, 48(10):1882-1891. WANG Y H, XU Y F, NIU J Q, et al.Preliminary studies on tissue expression profiles of Smad4 mRNA and its miRNAs in yak[J]. Acta Veterinaria et Zootechnica Sinica, 2017, 48(10):1882-1891.(in Chinese)
[12] 高真真.生长分化因子9及其信号通路蛋白(ALK5、Smad2和Smad4)在成年鹌鹑和鸡睾丸中的表达与定位[D].太谷:山西农业大学, 2014. GAO Z Z.Expression and localization of growth differentiation factor 9 and its signaling system proteins(ALK5, Smad2 and Smad4) in adult Japanese quail and chicken testes[D].Taigu:Shanxi Agricultural University, 2014.(in Chinese)
[13] 鄢胜飞, 张秀芳, 黄玥萌, 等.水牛Smad4基因的克隆及其真核表达载体的构建[J]. 中国畜牧兽医, 2017, 44(8):2369-2377. YAN S F, ZHANG X F, HUANG Y M, et al.Cloning and construction of eukaryotic expression vector of buffalo Smad4 gene[J]. China Animal Husbandry & Veterinary Medicine, 2017, 44(8):2369-2377.(in Chinese)
[14] 刘宇, 唐正严, 李冠军, 等. 人Smad4克隆及真核表达载体的构建与鉴定[J]. 临床泌尿外科杂志, 2009, 24(2):150-152. LIU Y, TANG Z Y, LI G J, et al.Clone of human Smad4 gene and construction of its eukaryotic expression vector[J]. Journal of Clinical Urology, 2009, 24(2):150-152.(in Chinese)
[15] 张小辉, 张路培, 许尚忠, 等.牛Smad4基因cDNA克隆及生物信息学分析[J]. 畜牧兽医学报, 2007, 38(4):321-325. ZHANG X H, ZHANG L P, XU S Z, et al.Cloning and bioinforinaton analysis of cDNA encoding cattle Smad4 gene[J]. Acta Veterinaria et Zootechnica Sinica, 2007, 38(4):321-325.(in Chinese)
[16] 赵永祥, 刘吉英, 潘增祥, 等.二花脸猪Smad4基因的克隆与卵巢组织mRNA的表达水平[J]. 中国农业科学, 2012, 45(23):4883-4890. ZHAO Y X, LIU J Y, PAN Z X, et al.Cloning and mRNA expression of Smad4 gene in ovaries of Erhualian pig[J]. Scientia Agricultura Sinica, 2012, 45(23):4883-4890.(in Chinese)
[17] 刘洪运, 韩香玉, 刘明晓, 等.梅花鹿Smad2与Smad4基因的克隆及其在鹿茸顶端组织的表达[J]. 黑龙江畜牧兽医, 2019, 3:6-10. LIU H Y, HAN X Y, LIU M X, et al.Cloning of Smad2 and Smad4 genes from sika deer and their expression in antler tissue[J]. Heilongjiang Animal Science and Veterinary Medicine, 2019, 3:6-10.(in Chinese)
[18] 张好放, 郁二蒙, 王广军, 等.草鱼TGF-β1、Smad4基因的克隆及真核过表达和RNA干扰表达载体的构建[J]. 中国农业大学学报, 2017, 22(4):120-128. ZHANG H F, YU E M, WANG G J, et al.Cloning of TGF-β1, Smad4 and construction of eukaryon and RNAi expression vector from Ctenopharyngodon idellus[J]. Journal of China Agricultural University, 2017, 22(4):120-128.(in Chinese)
[19] 吕池波, 郁二蒙, 谢骏, 等.草鱼Smad4基因的克隆、生物信息学分析及反义真核载体的构建[J]. 华南农业大学学报, 2013, 34(4):517-523. LYU C B, YU E M, XIE J, et al.Cloning of Smad4, bioinformatics analysis and construction of antisense smad4 gene expression vector from Ctenopharyngodon idellus[J]. Journal of South China Agricultural University, 2013, 34(4):517-523.(in Chinese)
[20] HUANG Z, YUAN X, WANG M, et al.Molecular cloning of the Smad4 gene and its mRNA expression analysis in ovarian follicles of the Yangzhou goose (Anser cygnoides)[J]. British Poultry Science, 2016, 57(4):515-521.
[21] UMAR S, STECHER G, LI M, et al.MegaX:Molecular evolutionary genetics analysis across computing platforms[J]. Molecular Biology and Evolution, 2018, 35(6):1547-1549.
[22] MCCARTHY A J, CHETTY R.Smad4/DPC4[J]. Journal of Clinical Pathology, 2018, 71(8):661.
[23] 李振芳.三角帆蚌Smads基因的表达及对创伤修复的探究[D].南昌:南昌大学, 2019. LI Z F.Expression of Smads genes in Hyiopsis cumingii and exploration of wound repair[D].Nanchang:Nanchang University, 2019.(in Chinese)
[24] 吐来力江·哈木太.MSTN/Smad信号通路与阿勒泰羊肌肉生长发育、产肉性能关联性的研究[D].乌鲁木齐:新疆农业大学, 2014. TULAILIJIANG H.MSTN/Smad signaling pathways and altai sheep muscle growthand comparative study of meat performance relevance[D].Urumqi:Xinjiang Agricultural University, 2014.(in Chinese)
[25] 黄菲菲, 张阳, 汪洋, 等.脂多糖刺激后不同时间点断奶仔猪回肠和结肠中转化生长因子β1及Smads信号通路的动态变化[J]. 中国畜牧杂志, 2020, 56(12):104-108. HUANG F F, ZHANG Y, WANG Y, et al.Developmental changes of TGF-β1 and smads signaling pathway in ileum and colon of piglets at different timepoints after LPS challenge[J]. Chinese Journal of Animal Science, 2020, 56(12):104-108.(in Chinese)
[26] 刘洪运.miRNA-18a和miRNA-106b对梅花鹿Smad家族调控作用的初步研究[D].长春:吉林农业大学, 2019. LIU H Y.Regulatory effects of miRNA-18a and miRNA-106b on Smad family of sika deer[D].Changchun:Jilin Agricultural University, 2019.(in Chinese)

Cloning, Bioinformatics and Tissue Expression Analysis of Smad4 Gene in Chicken (Gallus gallus)

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

References

Related Articles 15

Recommended Articles

Metrics

Comments

[1]	DU Yuanjun, DUAN Guochao, LIU Wei, LI Dapeng, HAN Haixia, LEI Qiuxia, ZHOU Yan, LIU Jie, WANG Jie, CAO Dingguo, LI Fuwei, CHEN Fu. Effect of Translucent on Egg Quality and Its Relationship with Intestinal Function of Laying Hen [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(6): 2305-2311.
[2]	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.
[3]	WANG Haorui, YU Baojun, YANG Jiayi, WEI Rong, FU Xi, WANG Chuanchuan, ZHANG Di, LI Desheng, CAI Zhengyun, GU Yaling, ZHANG Juan. Bioinformatics Analysis and Tissue Expression of DERA Gene in Jingyuan Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1764-1773.
[4]	LIU Jialong, ZHANG Yiwen, ZHANG Yueran, SUN Qijuan, CHEN Jian, HE Lei, JIA Yanyan, DING Ke, YU Zuhua. Cloning,Prokaryotic Expression and Bioinformatics Analysis of ELAVL1 Gene in Chicken [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1807-1817.
[5]	WU Jingxuan, SUN Zheng, MA Xuelian, ZHANG Liyang, LIAO Xiudong, LUO Xugang, LI Sufen, LYU Lin. A Survey on Magnesium Contents in Feed Resources for Livestock and Poultry in China [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 1888-1898.
[6]	JIANG Shanyi, ZHOU Bijun, WEN Ming, WANG Kaigong. Prevention of Traditional Chinese Medicine with Probiotics on Eimeria tenella [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2092-2102.
[7]	LIU Xin, CHEN Qun, QIU Yulang, LI Lin, HOU Guoxi, YAN Xiaogang, LI Zhonghe, CHEN Long, GAO Xingai, LIU Dongmei, ZHAI Di, YAN Qiuliang. Research Progress on the Production Mechanism and Control Technology of Chicken Manure Composting Nitrogen-containing and Sulfur-containing Waste Gases [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(5): 2166-2174.
[8]	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.
[9]	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.
[10]	WU Zhi, ZHANG Cong, YUAN Huisha, DONG Hongyan, DAI Lixia, JIA Luqi, WU Shuang, ZHU Shanyuan. Isolation,Identification,Virulence and Drug Resistance of Salmonella from Yellow Chicken [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(4): 1695-1706.
[11]	HUANG Xiaoqian, YUAN Junzhao, LIU Jinhe, ZOU Hui, GU Jianhong, YUAN Yan, LIU Xuezhong, LIU Zongping, BIAN Jianchun. Protective Effect of Honokiol on Cadmium-induced Renal Injury in Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 1007-1015.
[12]	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.
[13]	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.
[14]	LI Xinran, ZHANG Qingyan, ZHA Weiwei, ZHU Miao. Bioinformatics Analysis and Expression of Wnt Gene Family in Chickens [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 15-25.
[15]	XIE Jingjing, REN Mengting, FAN Jiahui, JIAO Yaru, HAN Zongxi, MA Deying. Study on Immune Response of Infectious Bronchitis Virus to Chickens and Antiviral Effect of Recombinant Avian Beta-defensins 11 and 12 [J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(1): 246-259.