山羊<i>KLF5</i>基因生物学特征与时空表达分析

doi:10.7668/hbnxb.20190562

摘要/Abstract

摘要： 为阐明KLF5在山羊不同组织和不同诱导分化阶段的肌内脂肪细胞中的表达特性，利用降落PCR克隆山羊KLF5基因cDNA序列，通过各种在线工具分析山羊KLF5基因的生物学特征，利用实时荧光定量PCR技术分析KLF5基因在山羊不同组织和不同诱导分化阶段的肌内脂肪细胞中的表达水平。结果表明，克隆获得山羊KLF5基因序列1 735 bp，其中包括1 365 bp完整的开放阅读框（ORF）、8 bp的5'非翻译区（UTR）和362 bp的3'UTR。蛋白预测显示，山羊KLF5编码454个氨基酸，合成不具有跨膜结构和信号肽的不稳定的亲水性蛋白。组织表达谱显示KLF5基因在山羊心、肝、脾、肺、肾、背最长肌、皮下脂肪和腹间脂肪等组织中均有表达，但在肺组织中的表达量较高，显著高于除腹间脂肪外的其他组织中的表达（P<0.05）。同时，细胞时序表达谱表明KLF5在分化的山羊肌内脂肪细胞中的表达水平均高于肌内前体脂肪细胞中的表达水平，且在诱导分化第5天时的表达量最高，显著高于第0天的表达量（P<0.05）。研究结果发现，具有3个锌指结构域等复杂结构的KLF5蛋白可能在山羊肌内脂肪细胞分化后期发挥正向的调控作用，并促进山羊肌内脂肪沉积。

关键词: 山羊, KLF5基因, 克隆, 生物学特征, 表达特性

Abstract: ln order to elucidate the expression patterns of KLF5 in goat different tissues and different differentiated stages of intramuscular adipocytes, the cloning of goat KLF5 was performed by Touchdown PCR technique, the biological characteristics of goat KLF5 were analyzed by a variety of online tools. In addition, the expression levels of goat KLF5 in different tissues and different differentiated stages of intramuscular adipocytes were investigated by qPCR technique. The obtained length of goat KLF5 sequence was 1 735 bp, including 1 365 bp complete ORF, 8 bp 5'UTR and 362 bp 3'UTR. Goat KLF5 gene encoded 454 amino acids and formed an unstable and hydrophobic protein without transmembrane domains and signal peptides. Tissue expression profile revealed that goat KLF5 gene was expressed in all the analyzed tissues including heart, liver, spleen, lung, kidney, longissimus dorsi, subcutaneous fat and abdominal fat, while its expression was the highest in the lung tissue being significantly higher than that the other analyzed tissues (P<0.05) except abdominal fat. Meanwhile, time-series expression profile indicated that the expression level of KLF5 in induced intramuscular adipocytes was higher than that in intramuscular preadipocytes, and the expression level at day 5 was highest and significantly higher than that at day 0(P< 0.05). It found that goat KLF5 protein with complex structures, such as three Zinc Finger domains etc., might play positive roles in the late differentiation of goat intramuscular adipocytes, and promoted goat intramuscular fat deposition.

Key words: Goat, KLF5 gene, Clone, Biological characteristic, Expression characteristics

中图分类号:

Q78
S826

谢光杰, 杜宇, 许晴, 马洁琼, 邱翔, 林亚秋. 山羊KLF5基因生物学特征与时空表达分析[J]. 华北农学报, 2020, 35(3): 225-232. doi: 10.7668/hbnxb.20190562.

XIE Guangjie, DU Yu, XU Qing, MA Jieqiong, QIU Xiang, LIN Yaqiu. Biological Characteristic and Spatio-Temporal Expression Analysis of KLF5 Gene in Goat[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2020, 35(3): 225-232. doi: 10.7668/hbnxb.20190562.

http://www.hbnxb.net/CN/Y2020/V35/I3/225

参考文献

[1] 谢伏瞻. 中国统计年鉴-2008[M]. 北京:中国统计出版社, 2008.
Xie F Z. China statistical yearbook-2008[M]. Beijing:China Statistics Press, 2008.
[2] 宁吉喆. 中国统计年鉴-2018[M]. 北京:中国统计出版社, 2018.
Ning J Z. China statistical yearbook-2018[M]. Beijing:China Statistics Press, 2018.
[3] Hocquette J F, Gondret F, Baéza E, Médale F, Jurie C, Pethick D W. Intramuscular fat content in meat-producing animals:development, genetic and nutritional control, and identification of putative markers[J]. Animal, 2010, 4(2):303-319. doi:10.1017/S1751731109991091.
[4] Pearson R, Fleetwood J, Eaton S, Crossley M, Bao S S. Krüppel-like transcription factors:A functional family[J]. The International Journal of Biochemistry & Cell Biology, 2008, 40(10):1996-2001. doi:10.1016/j.biocel.2007.07.018.
[5] McConnell B B, Yang V W. Mammalian Krüppel-like factors in health and diseases[J]. Physiological Reviews, 2010, 90(4):1337-1381. doi:10.1152/physrev.00058.2009.
[6] Pei J M, Grishin N V. A new family of predicted Krüppel-Like factor genes and pseudogenes in placental mammals[J]. PLoS One, 2013, 8(11):e81109. doi:10.1371/journal.pone.0081109.
[7] Yang C T, Ma R, Axton R A, Jackson M, Taylor A H, Fidanza A, Marenah L, Frayne J, Mountford J C, Forrester L M. Activation of KLF1 enhances the differentiation and maturation of red blood cells from human pluripotent stem cells[J]. Stem Cells, 2017, 35(4):886-897. doi:10.1002/stem.2562.
[8] Ghaleb A M, Elkarim E A, Bialkowska A B, Yang V W. KLF4 suppresses tumor formation in genetic and pharmacological mouse models of colonic tumorigenesis[J]. Molecular Cancer Research, 2016, 14(4):385-396. doi:10.1158/1541-7786.MCR-15-0410.
[9] Gao Y, Li H Z, Ma X, Fan Y,Ni D, Zhang Y, Huang Q B, Liu K, Li X T, Wang L, Gu L Y, Yao Y X, Ai Q, Du Q S, Song E L, Zhang X. KLF6 suppresses metastasis of clear cell renal cell carcinoma via transcriptional repression of E2F1[J]. Cancer Research, 2017, 77(2):330-342. doi:10.1158/0008-5472.CAN-16-0348.
[10] Dong J T, Chen C S. Essential role of KLF5 transcription factor in cell proliferation and differentiation and its implications for human diseases[J]. Cellular and Molecular Life Sciences, 2009, 66(16):2691-2706. doi:10.1007/s00018-009-0045-z.
[11] Okada H, Yamada M, Kamimoto K, Kok C Y, Kaneko K, Ema M, Miyajima A, Itoh T. The transcription factor Klf5 is essential for intrahepatic biliary epithelial tissue remodeling after cholestatic liver injury[J]. Journal of Biological Chemistry, 2018, 293(17):6214-6229. doi:10.1074/jbc.RA118.002372.
[12] Liu F F, Dong L, Yang X, Li D J, Shen Y Y, Liu Z L. KLF5 silence attenuates proliferation and epithelial-mesenchymal transition induction in Hep-2 cells through NF-κB signaling pathway[J]. European Review for Medical and Pharmacological Sciences, 2019, 23(9):3867-3875. doi:10.26355/eurrev_201905_17814.
[13] Li Y, Sui X N, Hu X Q, Hu Z. Overexpression of KLF5 inhibits puromycin-induced apoptosis of podocytes[J]. Molecular Medicine Reports, 2018,18(4):3843-3849. doi:10.3892/mmr.2018.9366.
[14] Oishi Y, Manabe I, Tobe K, Tsushima K, Shindo T, Fujiu K, Nishimura G, Maemura K, Yamauchi T, Kubota N, Suzuki R, Kitamura T, Akira S, Kadowaki T, Nagai R. Krüppel-like transcription factor KLF5 is a key regulator of adipocyte differentiation[J]. Cell Metabolism, 2005, 1(1):27-39. doi:10.1016/j.cmet.2004.11.005.
[15] Zhu S M, Cheng G, Zhu H L, Guan G C. A study of genes involved in adipocyte differentiation[J]. Journal of Pediatric Endocrinology and Metabolism, 2015, 28(1-2):93-99. doi:10.1515/jpem-2014-0002.
[16] Oishi Y, Manabe I, Tobe K, Ohsugi M, Kubota T, Fujiu K, Maemura K, Kubota N, Kadowaki T, Nagai R. SUMOylation of Krüppel-like transcription factor 5 acts as a molecular switch in transcriptional programs of lipid metabolism involving PPAR-δ[J]. Nature Medicine, 2008, 14(6):656-666. doi:10.1038/nm1756.
[17] Bonnet M, Bernard L, Bes S, Leroux C. Selection of reference genes for quantitative Real-time PCR normalisation in adipose tissue, muscle, liver and mammary gland from ruminants[J]. Animal, 2013, 7(8):1344-1353. doi:10.1017/S1751731113000475.
[18] Ardito F, Giuliani M, Perrone D, Troiano G, Muzio L L. The crucial role of protein phosphorylation in cell signaling and its use as targeted therapy (Review)[J]. International Journal of Molecular Medicine, 2017, 40(2):271-280. doi:10.3892/ijmm.2017.3036.
[19] 赵新元, 沈丙权, 秦伟捷, 钱小红. 基于生物质谱的蛋白质O-GalNAc糖基化修饰研究进展[J]. 中国科学, 2018, 48(2):113-123. doi:10.1360/N052017-00137.
Zhao X Y, Shen B Q, Qin W J, Qian X H. Advances in O-GalNAc glycosylation research based on biological mass spectrometry[J]. Scientia Sinica Vitae, 2018, 48(2):113-123.
[20] Spiro R G. Protein glycosylation:nature, distribution, enzymatic formation, and disease implications of glycopeptide bonds[J]. Glycobiology, 2002, 12(4):43-56. doi:10.1093/glycob/12.4.43R.
[21] Garton M, Najafabadi H S, Schmitges F W, Radovani E, Hughes T R, Kim P M. A structural approach reveals how neighbouring C2H2 zinc fingers influence DNA binding specificity[J]. Nucleic Acids Research, 2015, 43(19):9147-9157. doi:10.1093/nar/gkv919.
[22] Wan H J, Luo F M, Wert S E, Zhang L Q, Xu Y, Ikegami M, Maeda Y, Bell S M, Whitsett J A. Kruppel-like factor 5 is required for perinatal lung morphogenesis and function[J]. Development, 2008, 135(15):2563-2572. doi:10.1242/dev.021964.
[23] Zhang X X, Lian T, Ran J S, Li Z Q, Han S S, Liu Y P. KLF5 functions in proliferation, differentiation, and apoptosis of chicken satellite cells[J]. 3 Biotech, 2019, 9(6):222. doi:10.1007/s13205-019-1752-2.

选择文件类型/文献管理软件名称

选择包含的内容

山羊KLF5基因生物学特征与时空表达分析

Biological Characteristic and Spatio-Temporal Expression Analysis of KLF5 Gene in Goat

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价

[1]	金一锋, 高岩松, 王琦, 王萌萌, 赵迪, 熊毅, 陈阳. *草地早熟禾蛋白激酶SnRK2.4* 基因克隆及非生物胁迫响应分析**[J]. 华北农学报, 2022, 37(5): 9-15.
[2]	张冬梅, 冯亚艳, 修志君, 杨春芳, 杜美娥, 李得宙, 张笑宇. *硅酸钠诱导的马铃薯抗黑痣病StWRKY11基因克隆及生物信息学分析*[J]. 华北农学报, 2022, 37(5): 194-203.
[3]	郑玉斯, 王培, 郭颖, 白丽蓉, 喻达辉, 赵森. *合浦珠母贝PfCLEC17A* 基因的克隆及表达分析**[J]. 华北农学报, 2022, 37(5): 230-238.
[4]	肖士奎, 李芳, 张文婷, 吕淑芳, 史国安, 吴疆, 范丙友. *芍药ACS基因的克隆及其蛋白质原核表达*[J]. 华北农学报, 2022, 37(5): 36-44.
[5]	侯瑞泽, 侯玉翔, 许小勇, 李璿, 李梅兰. 普通白菜CYP同源基因克隆与表达分析[J]. 华北农学报, 2022, 37(5): 45-51.
[6]	吉祥, 宋志成, 韦晓玲, 杨煜, 隋炯明, 郭宝太. 马铃薯卷叶病毒与S病毒重组双CP多克隆抗体的制备[J]. 华北农学报, 2022, 37(4): 212-219.
[7]	邓婕, 王瀚林, 李有玉, 王自娥, 陈晓华, 刘迪秋. 岷江百合多聚半乳糖醛酸酶抑制蛋白基因及其启动子的克隆与分析[J]. 华北农学报, 2022, 37(4): 45-52.
[8]	王亚男, 梁岩岩, 严文培, 张银萍, 李强, 吴秀菊. *北细辛AhOMT1基因的克隆及原核表达分析*[J]. 华北农学报, 2022, 37(4): 62-70.
[9]	张晋玉, 徐新娟, 晁毛妮, 张晓红, 吴向远, 高际涛, 黄中文. *大豆GmZAT12基因的克隆和表达分析*[J]. 华北农学报, 2022, 37(3): 1-7.
[10]	贾志强, 许云玉, 高雪, 陶宏征, 陈增敏, 刘雅婷, 李永忠. *辣椒CaWRKY30转录因子的克隆、亚细胞定位及番茄斑萎病毒侵染下的表达分析*[J]. 华北农学报, 2022, 37(3): 186-192.
[11]	吉颖, 孙枫, 吴淑华, 李硕, 涂丽琴, 高丹娜, 崔晓艳, 陈新, 季英华, 郭青云. 江苏大豆上分离的豇豆轻斑驳病毒基因组序列克隆及特征分析[J]. 华北农学报, 2022, 37(3): 200-205.
[12]	安清明, 王星, 吴震洋, 孟金柱, 赵园园, 宋兴超. 基于转录组挖掘不同性别贵州白山羊肌肉生长发育的关键基因及PI3K/ATK信号通路分析[J]. 华北农学报, 2022, 37(3): 213-222.
[13]	冉黎, 吕锦诗, 张浩, 王永, 朱江江, 李艳艳, 孟庆勇, 林亚秋. *过表达山羊APOC3基因促进肌内脂肪细胞分化*[J]. 华北农学报, 2022, 37(3): 223-230.
[14]	程春红, 游经番, 蔡兆明, 叶春宏, 陈丽. *茎瘤芥BjuEAR1-1的克隆与表达分析*[J]. 华北农学报, 2022, 37(3): 37-43.
[15]	唐忠丽, 逯晓楠, 赵蕊, 刘庆华, 李梅兰, 雷逢进, 许小勇. *黄皮西葫芦类胡萝卜素合成酶基因的鉴定及PSY1和LCYE2克隆分析*[J]. 华北农学报, 2022, 37(3): 60-67.

模态框（Modal）标题

选择文件类型/文献管理软件名称

选择包含的内容

山羊KLF5基因生物学特征与时空表达分析

Biological Characteristic and Spatio-Temporal Expression Analysis of KLF5 Gene in Goat

PDF

可视化

被引次数

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价