牦牛KLF5、KLF6、KLF7基因的克隆表达及其与肌内脂肪含量的相关性分析

doi:10.11843/j.issn.0366-6964.2017.03.004

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (3): 416-424.doi: 10.11843/j.issn.0366-6964.2017.03.004

牦牛KLF5、KLF6、KLF7基因的克隆表达及其与肌内脂肪含量的相关性分析

朱江江^1,2，林亚秋^1,2，左璐璐¹，王永^1,2，柏雪^2*，江明锋²

(1. 青藏高原动物遗传资源保护与利用四川省重点实验室，成都 610041；2. 动物遗传育种学国家民委-教育部重点实验室,成都 610041)

收稿日期:2016-09-12 出版日期:2017-03-23 发布日期:2017-03-23
通讯作者: 柏雪，硕士，讲师，主要从事动物营养学研究，E-mail: yukivy@163.com
作者简介:朱江江（1986-），男，湖北钟祥人，博士，助理研究员，主要从事动物遗传育种学研究，E-mail: zhujiang4656@hotmail.com
基金资助:
四川省青年科技创新团队项目(2015TD0025)

Molecular Cloning, Tissue Expression of KLF5, KLF6, KLF7 Genes, and Their Correlations with Intramuscular Fat Content in Yak

ZHU Jiang-jiang^1,2, LIN Ya-qiu^1,2, ZUO Lu-lu¹, WANG Yong^1,2, BAI Xue^2* , JIANG Ming-feng²

(1. Key Laboratory of Sichuan Province for Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation, Chengdu 610041, China; 2. Key Laboratory of State Ethnic Affairs Commission and Ministry of Education for Animal Genetics & Breeding, Chengdu 610041, China)

Received:2016-09-12 Online:2017-03-23 Published:2017-03-23

摘要/Abstract

摘要：

本研究旨在克隆牦牛Krüppel样因子（KLF5、KLF6、KLF7）的CDS区序列，鉴定其在牦牛不同组织中的表达水平，并分析其表达水平与牦牛肌内脂肪（IMF）的相关性，从而为研究KLFs在牦牛脂质代谢中的功能提供试验依据。选取4~6周岁的健康麦洼牦牛公牛7头，清晨空腹屠宰后，迅速采集脾、肺、肾、皮下脂肪和背最长肌组织样品，提取组织总RNA，利用RT-PCR法克隆牦牛KLF5、KLF6、KLF7基因序列，进行生物信息学分析，利用实时定量PCR法检测其mRNA的表达水平，并使用皮尔逊相关系数进行KLF5、KLF6、KLF7基因的表达量与IMF含量的相关性分析。结果表明，KLF5、KLF6、KLF7基因CDS全长分别为1 086、855和909 bp，分别编码361、284和302个氨基酸残基，且均与牛具有最近的亲缘关系，其次为山羊、绵羊、猪，而与鸡的亲缘关系较远。KLF5、KLF6、KLF7均在肺中具有较高的表达水平，而在背最长肌中的表达量较低，且与牦牛IMF含量均没有显著相关性。但KLF5的表达水平与KLF6、KLF7均具有显著的正相关性。这些结果为研究KLF5、KLF6、KLF7的功能提供了基础数据，也为揭示牦牛脂质代谢的调控机制提供了试验参考。

Abstract:

The aims of the study were to clone the CDS of krüppel-like factors (KLF5, KLF6, KLF7), determine their expression in various tissues, and analyze their correlation with intramuscular fat (IMF), which might provide basis for the functional research on KLFs in regulating lipid metabolism in yak. Seven healthy, 4-6 years old male Maiwa yaks were selected as experiment animals. After fasting slaughter, the tissue samples of spleen, lung, kidney, subcutaneous fat and longissimus dorsi muscle were collected for the total RNA extraction. The sequences of KLF5, KLF6 and KLF7 genes were cloned by RT-PCR, and analyzed using bioinformatics, respectively. The mRNA expression of KLFs were determined using real-time fluorescent quantitative PCR (qPCR). Pearson correlation coefficient was used for analyzing the correlation between the mRNA expression of KLFs and IMF content in yak. The CDS of KLF5, KLF6 and KLF7 were 1 086, 855 and 909 bp, encoding 361, 284 and 302 amino acids, respectively. The yak KLFs sequences were closest associated with bovine, followed by goat, sheep, pig, and more distant with chicken. The tissue expression of the KLF5, KLF6 and KLF7 were higher in lung, and lower in longissimus dorsi muscle. The expression of KLF5 was significantly correlated with the expression of KLF6 and KLF7, respectively. However, all of mRNA expression of KLFs were not correlated with IMF content in longissimus dorsi muscle of yak. These data provided basic information for the functional research of KLF5, KLF6, KLF7, and would be beneficial for understanding the regulatory mechanism of lipid metabolism in yak.

中图分类号:

朱江江，林亚秋，左璐璐，王永，柏雪，江明锋. 牦牛KLF5、KLF6、KLF7基因的克隆表达及其与肌内脂肪含量的相关性分析[J]. 畜牧兽医学报, 2017, 48(3): 416-424.

ZHU Jiang-jiang, LIN Ya-qiu, ZUO Lu-lu, WANG Yong, BAI Xue, JIANG Ming-feng. Molecular Cloning, Tissue Expression of KLF5, KLF6, KLF7 Genes, and Their Correlations with Intramuscular Fat Content in Yak[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(3): 416-424.

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参考文献

［1］熊倩, 张昭军, 方向东. KLFs对珠蛋白基因表达和红系分化的调控作用［J］. 中国生物化学与分子生物学报, 2011, 27(8): 693-699.
XIONG Q, ZHANG Z J, FANG X D. Regulation of KLFs on globin expression and erythroid differentiation［J］. Chinese Journal of Biochemistry and Molecular Biology, 2011, 27(8):693-699. (in Chinese)
［2］SUSKE G, BRUFORD E, PHILIPSEN S. Mammalian SP/KLF transcription factors: bring in the family ［J］. Genomics, 2005, 85(5): 551-556.
［3］SHINDO T, MANABE I, FUKUSHIMA Y, et al. Kruppel-like zinc-finger transcription factor KLF5/BTEB2 is a target for angiotensin II signaling and an essential regulator of cardiovascular remodeling［J］. Nat Med, 2002, 8(8): 856-863.
［4］陈林波, 张新军, 郭俊明. Krüppel样因子5的生物学功能及其与肿瘤发生的关系［J］. 中国细胞生物学学报, 2016, 38(5):647-656.
CHEN L B, ZHANG X J, GUO J M. The biological function of krüppel-like factor 5 and its association with cancer occurrence［J］. Chinese Journal of Cell Biology, 2016, 38(5): 647-656. (in Chinese)
［5］KREMER-TAL S, NARLA G, CHEN Y, et al. Downregulation of KLF6 is an early event in hepatocarcinogenesis, and stimulates proliferation while reducing differentiation［J］. J Hepatol, 2007, 46(4): 645-654.
［6］KANAZAWA A, KAWAMURA Y, SEKINE A, et al. Single nucleotide polymorphisms in the gene encoding Krüppel-like factor 7 are associated with type 2 diabetes［J］. Diabetologia, 2005, 48(7): 1315-1322.
［7］ZOBEL D P, ANDREASEN C H, BURGDORF K S, et al. Variation in the gene encoding Krüppel-like factor 7 influences body fat: studies of 14 818 Danes［J］. European J Endocrinol, 2009, 160(4): 603-609.
［8］RAY K. NASH: KLF6 activates PPARalpha signalling in hepatic steatosis［J］. Nat Rev Gastroenterol Hepatol, 2013, 10(3): 128.
［9］WEI S, ZHANG L, ZHOU X, et al. Emerging roles of zinc finger proteins in regulating adipogenesis［J］. Cell Mol Life Sci, 2013, 70(23): 4569-4584.
［10］OISHI Y, MANABE I, TOBE K, et al. Krüppel-like transcription factor KLF5 is a key regulator of adipocyte differentiation［J］. Cell Metab, 2005, 1(1): 27-39.
［11］SHI H B, ZHAO W S, LUO J, et al. Peroxisome proliferator-activated receptor gamma1 and gamma2 isoforms alter lipogenic gene networks in goat mammary epithelial cells to different extents［J］. J Dairy Sci, 2014, 97(9): 5437-5447.
［12］LEFTEROVA M I, LAZAR M A. New developments in adipogenesis［J］. Trends Endocrinol Metab, 2009, 20(3): 107-114.
［13］DROSATOS K, DROSATOS-TAMPAKAKI Z, KHAN R, et al. Inhibition of c-Jun-N-terminal kinase increases cardiac peroxisome proliferator-activated receptor alpha expression and fatty acid oxidation and prevents lipopolysaccharide-induced heart dysfunction［J］. J Biol Chem, 2011, 286(42): 36331-36339.
［14］ROE N D, STANDAGE S W, TIAN R. The relationship between KLF5 and PPARalpha in the heart: It's complicated［J］. Circ Res, 2016, 118(2): 193-195.
［15］CHAKRAVARTHY M V, LODHI I J, YIN L, et al. Identification of a physiologically relevant endogenous ligand for PPARalpha in liver［J］. Cell, 2009, 138(3): 476-488.
［16］SMAS C M, SUL H S. Pref-1, a protein containing EGF-like repeats, inhibits adipocyte differentiation［J］. Cell, 1993, 73(4): 725-734.
［17］LI D, YEA S, LI S, et al. Krüppel-like factor-6 promotes preadipocyte differentiation through histone deacetylase 3-dependent repression of DLK1［J］. J Biol Chem, 2005, 280(29): 26941-26952.
［18］BECHMANN L P, VETTER D, ISHIDA J, et al. Post-transcriptional activation of PPAR alpha by KLF6 in hepatic steatosis［J］. J Hepatol, 2013, 58(5): 1000-1006.
［19］ZHANG Z, WANG H, SUN Y, et al. Klf7 modulates the differentiation and proliferation of chicken preadipocyte［J］. Acta Biochim Biophys Sin (Shanghai), 2013, 45(4): 280-288.
［20］LAUB F, ALDABE R, FRIEDRICH V Jr, et al. Developmental expression of mouse krüppel-like transcription factor KLF7 suggests a potential role in neurogenesis［J］. Dev Biol, 2001, 233(2): 305-318.
［21］KAWAMURA Y, TANAKA Y, KAWAMORI R, et al. Overexpression of Krüppel-like factor 7 regulates adipocytokine gene expressions in human adipocytes and inhibits glucose-induced insulin secretion in pancreatic beta-cell line［J］. Mol Endocrinol, 2006, 20(4): 844-856.
［22］朱武政. 山羊脂肪沉积相关基因的表达及与肌内脂肪含量的相关性研究［D］. 成都：西南民族大学，2015.
ZHU W Z. Study on expression of genes related to fat deposition and association with intramuscular fat content［D］. Chengdu: Southwest University for Nationalities, 2015. (in Chinese)
［23］孙雨婷, 罗军, 朱江江, 等. 奶山羊短链脂肪酸受体GPR41基因的克隆及组织表达分析［J］. 畜牧兽医学报, 2012, 43(2): 319-323.
SUN Y T, LUO J, ZHU J J, et al. Cloning and analysis on tissue expression of GPR41 gene in dairy goat［J］. Acta Veterinaria et Zootechnica Sinica, 2012, 43(2):319-323. (in Chinese)
［24］朱武政, 林亚秋, 江明锋, 等. 肉用山羊脂代谢相关基因与肌内脂肪含量的相关性分析［J］. 畜牧兽医学报, 2016, 47(7):1333-1341.
ZHU W Z, LIN Y Q, JIANG M F, et al. Association analysis of intramuscular fat content and the expression of genes related to lipid metabolism in meat goat［J］. Acta Veterinaria et Zootechnica Sinica, 2016, 47(7):1333-1341. (in Chinese)
［25］罗晓林, 李洪泉, 文勇立, 等. 麦洼牦牛高产奶性能新品系(品种) 选育方案［J］. 黑龙江畜牧兽医,2015,（4):195-198.
LUO X L, LI H Q, WEN Y L, et al. A scheme of seletive breeding for a new line (variety) of Maiwa yak with high milk performance［J］. Heilongjang Animal Science and Veterinary Medicine, 2015,(4): 195-198. (in Chinese)
［26］LIU H N, REN F Z, JIANG L, et al. Fatty acid profile of yak milk from the Qinghai-Tibetan Plateau in different seasons and for different parities［J］. J Dairy Sci, 2011, 94(4):1724-1731.
［27］DONG S, LONG R, KANG M. Milking performance of China yak (Bos grunniens): A preliminary report［J］. Afr J Agr Res, 2007, 2(3): 52-57.
［28］周明亮, 蒋世海, 谢荣清, 等. 麦洼牦牛产奶变化规律研究［J］. 湖北农业科学, 2013, 52(19): 4730-4732.
ZHOU M L, JIANG S H, XIE R Q, et al. Study on the milk production pattern of Maiwa yak［J］. Hubei Agricultural Science, 2013, 52(19): 4730-4732. (in Chinese)
［29］CHEN Z, LEI T, CHEN X, et al. Porcine KLF gene family: Structure, mapping, and phylogenetic analysis［J］. Genomics, 2010, 95(2): 111-119.
［30］WANG W, LUO J, ZHONG Y, et al. Goat liver X receptor α, molecular cloning, functional characterization and regulating fatty acid synthesis in epithelial cells of goat mammary glands［J］. Gene, 2012, 505(1): 114-120.

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牦牛KLF5、KLF6、KLF7基因的克隆表达及其与肌内脂肪含量的相关性分析

Molecular Cloning, Tissue Expression of KLF5, KLF6, KLF7 Genes, and Their Correlations with Intramuscular Fat Content in Yak

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