贵州从江香猪PDK4、FGF10基因的克隆及组织表达分析

doi:10.16431/j.cnki.1671-7236.2017.12.002

《中国畜牧兽医》 ›› 2017, Vol. 44 ›› Issue (12): 3401-3409.doi: 10.16431/j.cnki.1671-7236.2017.12.002

贵州从江香猪PDK4、FGF10基因的克隆及组织表达分析

杨洋^1,2,3, 许厚强^1,2,3, 陈伟^1,2,4, 周迪^1,2,4, 徐敏^1,2,3, 张青青^1,2,3, 赵焕平^1,2,3, 孙成娟^1,2,3, 王圆圆^1,2,3, 张鸣^1,2,3, 杨涛^1,2,3

1. 贵州大学高原山地动物遗传育种与繁殖省部共建教育部重点实验室, 贵阳 550025;
2. 贵州省动物遗传育种与繁殖重点实验室, 贵阳 550025;
3. 贵州大学动物科学学院, 贵阳 550025;
4. 贵州大学生命科学学院, 贵阳 550025

收稿日期:2017-06-12 出版日期:2017-12-20 发布日期:2017-12-20
通讯作者: 许厚强 E-mail:gzdxxhq@163.com
作者简介:杨洋(1993-),男,安徽阜阳人,硕士生,研究方向:动物遗传育种与种质资源创新,E-mail:1037856957@qq.com
基金资助:
国家科技支撑计划（2015BAD03B02-3）；黔科合重大专项（黔科合NY字[2013]6008号）

Cloning and Tissue Expression Analysis of PDK4 and FGF10 Genes in Guizhou Congjiang Xiang Pig

YANG Yang^1,2,3, XU Hou-qiang^1,2,3, CHEN Wei^1,2,4, ZHOU Di^1,2,4, XU Min^1,2,3, ZHANG Qing-qing^1,2,3, ZHAO Huan-ping^1,2,3, SUN Cheng-juan^1,2,3, WANG Yuan-yuan^1,2,3, ZHANG Ming^1,2,3, YANG Tao^1,2,3

1. Key Laboratory of Animal Genetics, Breeding and Reproduction in the Platean Mountainous Region, Ministry of Education, Guizhou University, Guiyang 550025, China;
2. Key Laboratory of Animal Genetics, Breeding and Reproduction in Guizhou, Guiyang 550025, China;
3. College of Animal Science, Guizhou University, Guiyang 550025, China;
4. College of Life Science, Guizhou University, Guiyang 550025, China

Received:2017-06-12 Online:2017-12-20 Published:2017-12-20

摘要/Abstract

摘要：

试验旨在克隆获得PDK4、FGF10基因，并研究大白猪与从江香猪不同组织中PDK4、FGF10基因mRNA的表达差异。采用RT-PCR分别克隆从江香猪PDK4、FGF10基因并进行生物信息学分析，利用实时荧光定量PCR技术检测PDK4、FGF10基因在大白猪和从江香猪不同组织中mRNA的相对表达量。结果显示，从江香猪PDK4基因的编码区全长1 224 bp，编码407个氨基酸；FGF10基因的编码区全长636 bp，编码211个氨基酸。经BLAST软件进行同源性比对，发现从江香猪PDK4基因与羊、马、人的核苷酸序列同源性分别为93%、92%和91%；FGF10基因与羊、牛、人、鼠的核苷酸序列同源性分别为94%、93%、93%和90%。由PDK4基因系统进化树可知，从江香猪与牛、绵羊亲缘关系较近；由FGF10基因系统进化树可知，从江香猪与绵羊、牛、人、猕猴亲缘关系较近，与小鼠和鸡亲缘关系较远。实时荧光定量PCR结果显示，在从江香猪不同组织中，PDK4基因在肾脏中的表达量最高，在胃和脂肪中表达量较高，FGF10基因在胃中表达量最高，在肾脏和脂肪中表达量较高，两个基因在背最长肌中的表达量均最低；在大白猪的不同组织中，PDK4、FGF10基因在脂肪中的表达量均最高，PDK4基因在背最长肌中的表达量最低，而FGF10基因在心脏中表达量最低。本试验成功克隆了从江香猪PDK4、FGF10基因，并检测了其在大白猪与从江香猪不同组织中的表达，为进一步研究PDK4、FGF10基因在脂质代谢及脂肪沉积等方面的调控作用提供科学依据。

关键词: 从江香猪; PDK4基因; FGF10基因; 实时荧光定量PCR

Abstract:

The objective of this study was to clone PDK4 and FGF10 genes, and investigate the expression level of PDK4 and FGF10 genes mRNA in different tissues of Large White pig and Congjiang Xiang pig. The PDK4 and FGF10 genes were cloned by RT-PCR and analyzed by bioinformatics, the relative expression of PDK4 and FGF10 genes were detected by Real-time PCR. The results showed that the coding region of PDK4 gene was 1 224 bp, encoding 407 amino acids; The coding region of FGF10 gene was 636 bp and encoded 211 amino acids. The homologies of nucleotide sequences of PDK4 gene with sheep, horse and human were 93%, 92% and 91%,respectively. The homologies of nucleotide sequences of FGF10 gene with sheep, cattle, human and mouse were 94%,93%, 93% and 90%, respectively. The phylogenetic tree of PDK4 gene showed that the genetic relationship of Congjiang Xiang pig, cattle and sheep were very close, the phylogenetic tree of FGF10 gene indicated that the genetic relationship of Congjiang Xiang pig, cattle, sheep, human and macaque were very close, but the genetic relationship of Congjiang Xiang pig, rat and chicken were far away. Real-time PCR results showed that, in different tissues of Congjiang Xiang pig,PDK4 gene expression in kidney tissue was higher than other tissues, with a higher expression in stomach and adipose as well,FGF10 gene expression in stomach tissue was higher than other tissues, with a higher expression in kidney and adipose as well, but both of PDK4 and FGF10 genes expression were the lowest in longissimus dorsi. In different tissues of Large White pig, both of PDK4 and FGF10 genes were expressed the highest in adipose than other tissues, PDK4 gene expression in longissimus dorsi was the lowest, while the FGF10 gene expression the lowest in heart. This study successfully cloned the PDK4 and FGF10 genes of Large White pig and Congjiang Xiang pig,and detected the relative expression of PDK4 and FGF10 genes in different tissues of Large White pig and Congjiang Xiang pig, and also provided scientific basis for further study on regulation of PDK4 and FGF10 genes on lipid metabolism and deposition.

Key words: Congjiang Xiang pig; PDK4 gene; FGF10 gene; Real-time PCR

中图分类号:

S828

杨洋, 许厚强, 陈伟, 周迪, 徐敏, 张青青, 赵焕平, 孙成娟, 王圆圆, 张鸣, 杨涛. 贵州从江香猪PDK4、FGF10基因的克隆及组织表达分析[J]. 《中国畜牧兽医》, 2017, 44(12): 3401-3409.

YANG Yang, XU Hou-qiang, CHEN Wei, ZHOU Di, XU Min, ZHANG Qing-qing, ZHAO Huan-ping, SUN Cheng-juan, WANG Yuan-yuan, ZHANG Ming, YANG Tao. Cloning and Tissue Expression Analysis of PDK4 and FGF10 Genes in Guizhou Congjiang Xiang Pig[J]. , 2017, 44(12): 3401-3409.

参考文献

[1] Sugden M C, Holness M J. Mechanisms underlying regulation of the expression and activities of the mammalian pyruvate dehydrogenase kinases[J]. Archives of Physiology and Biochemistry, 2006, 112(3):139.
[2] Dixit D, Ahmad F, Ghildiyal R, et al. CK2 inhibition induced PDK4-AMPK axis regulates metabolic adaptation and survival responses in glioma[J]. Experimental Cell Research, 2016, 344(1):132.
[3] Aminizadeh S, Habibi A, Marefati H, et al. The role of pyruvate dehydrogenase kinase 4(PDK4) on the expression of citrate synthase in the skeletal muscle after 4 weeks of endurance training in male wistar rats[J]. Rafsanjan University of Medical Sciences, 2017,16(3):191-202.
[4] 奚子英, 何小梅, 陈刚,等.鸡PDK4基因的克隆及组织差异表达研究[J]. 中国家禽, 2015, 37(15):6-10.
[5] 杜姗姗,周文婷,高亚可,等. 水牛FGF10基因的克隆分析及其在不同组织中表达情况的研究[J]. 中国畜牧杂志, 2016, 52(15):15-19.
[6] Thisse B, Thisse C. Functions and regulations of fibroblast growth factor signaling during embryonic development[J]. Developmental Biology, 2005, 287(2):390-402.
[7] 刘玉芳, 康晓龙, 方美英.滩羊FGF21基因CDS区克隆表达及生物信息学分析[J]. 中国畜牧杂志, 2016,52(13):23-28.
[8] Itoh N. FGFs as multifunctional signaling molecules:Diversity of structure and function[J]. Seikagaku, 2001, 73(7):525-535.
[9] Ohta H, Konishi M, Itoh N. FGF10 and FGF21 as regulators in adipocyte development and metabolism[J]. Endocrine Metabolic and Immune Disorders Drug Targets, 2011, 11(11):302-309.
[10] Ohta H, Itoh N. Roles of FGFs as adipokines in adipose tissue development, remodeling, and metabolism[J]. Frontiers in Endocrinology, 2014, 5:18.
[11] Regatieri I C, Pereira G L, Neto A R T, et al. Polymorphisms in MCT1, CD147, PDK4 and DMRT3 genes in Arabian and Quarter horses[J]. Journal of Equine Veterinary Science, 2016, 344(1):132.
[12] 徐凯,王效京,刘宏,等.过氧化物酶体增殖物激活受体α在猪肝脏组织中发育性表达研究[J]. 中国畜牧兽医,2014, 41(10):178-182.
[13] 周招洪,陈代文,田刚,等.过氧化物酶体增殖物激活受体γ辅激活因子1α与肌纤维类型转化及其表达调控研究进展[J]. 中国畜牧兽医,2015, 42(10):2636-2643.
[14] Degenhardt T, Saranarki A, Malinen M, et al. Three members of the human pyruvate dehydrogenase kinase gene family are direct targets of the peroxisome proliferator-activated receptor beta/delta[J]. Journal of Molecular Biology, 2007, 372(2):341-355.
[15] 张罕星,朱晓彤,束刚,等.猪肌内脂肪前体细胞与皮下脂肪前体细胞分化过程中基因差异表达分析[J]. 中国农业科学,2008, 41(11):3760-3768.
[16] 田家伟.猪PDK4基因启动子的克隆及其活性分析[D]. 重庆:西南大学, 2012.
[17] 张榕婧, 何小梅,闫景旭,等. 鸡PDK4基因的组织表达分析[J]. 广东农业科学, 2014, 41(15):121-125.
[18] Yamasaki M, Miyake A, Tagashira S, et al. Structure and expression of the rat mRNA encoding a novel member of the fibroblast growth factor family[J]. Journal of Biological Chemistry, 1996, 271(27):15918-15921.
[19] Yamasaki M, Emoto H, Konishi M, et al. FGF-10 is a growth factor for preadipocytes in white adipose tissue[J]. Biochemical and Biophysical Research Communications, 1999, 258(1):109-112.
[20] 李倩,林亚秋,朱江江,等. 山羊FGF10基因的克隆及其表达分析[J]. 农业生物技术学报, 2016, 24(11):1726-1733.
[21] Konishi M, Asaki T, Koike N, et al. Role of FGF10 in cell proliferation in white adipose tissue[J]. Molecular and Cellular Endocrinology, 2006, 249(1-2):71-77.
[22] 张迅轶.重组人成纤维细胞生长因子10间充质干细胞诱导人脂肪干细胞分化的研究[D].上海:第二军医大学, 2007.

贵州从江香猪PDK4、FGF10基因的克隆及组织表达分析

Cloning and Tissue Expression Analysis of PDK4 and FGF10 Genes in Guizhou Congjiang Xiang Pig

PDF

可视化

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]	麻宝艺, 盛陈艳, 刘宏莹, 马青霞, 汪婷婷, 李健明, 时坤, 杜锐, 冷雪. BVDV 1型和2型TaqMan双重实时荧光定量PCR检测方法的建立[J]. 中国畜牧兽医, 2022, 49(6): 2326-2335.
[2]	吕晓楠, 徐立蒲, 张文, 王娜, 曹欢, 王小亮, 王姝, 王静波. 感染鲤浮肿病毒镜鲤的组织病理变化及病毒分布规律研究[J]. 中国畜牧兽医, 2022, 49(3): 1077-1084.
[3]	张军芳, 孙建富, 孙斌, 唐琳, 王英, 王恩泽, 崔岩, 李强, 严昌国, 李香子. 延边牛PPARγ基因的克隆及生物信息学分析[J]. 中国畜牧兽医, 2021, 48(8): 2695-2704.
[4]	苗曼宁, 郭益文, 胡德宝, 曾雨晗, 李新, 张林林, 丁向彬, 郭宏. MSTN基因编辑牛肌肉转录组测序及生物信息学分析[J]. 中国畜牧兽医, 2021, 48(7): 2271-2281.
[5]	李恒, 字向东. 牦牛跨膜附睾蛋白1基因的克隆及其在卵丘卵母细胞复合体中的表达分析[J]. 中国畜牧兽医, 2021, 48(4): 1284-1293.
[6]	李建梅, 季正剑, 刘梅, 沈欣悦, 程旭, 俞燕, 戴亚斌. 藏鸡和隐性白羽鸡感染柔嫩艾美耳球虫后免疫相关基因的表达变化分析[J]. 中国畜牧兽医, 2021, 48(4): 1440-1448.
[7]	陈磊, 袁枫, 贺三刚, 玛依拉, 李文蓉. 绵羊FGF10基因克隆、序列分析及其在毛囊发育中的表达分析[J]. 中国畜牧兽医, 2021, 48(2): 425-432.
[8]	戴政列, 朱静静, 陈振宇, 周晓龙, 汪涵, 李向臣, 赵阿勇, 杨松柏. 猪SENP1基因克隆、生物信息学分析与表达研究[J]. 中国畜牧兽医, 2021, 48(12): 4382-4390.
[9]	姜懿轩, 王亚玲, 邢珊珊, 宋国华, 许会芬, 李明. 新西兰白兔MSTN基因克隆、生物信息学分析及表达谱构建[J]. 中国畜牧兽医, 2021, 48(10): 3501-3511.
[10]	李志强, 陶晨雨, 魏奥龙, 贾青. 猪性控精子纯度实时荧光定量PCR检测方法的建立[J]. 中国畜牧兽医, 2021, 48(10): 3682-3690.
[11]	王征帆, 朱利塞, 王娟, 李祎云, 向蕊, 应碧云, 王贵平, 贾爱卿, 白挨泉. PDCoV与TGEV双重实时荧光定量PCR检测方法的建立及初步应用[J]. 中国畜牧兽医, 2021, 48(10): 3855-3863.
[12]	李安奇, 蒋蕾, 苏晓彤, 昝林森, 王洪宝. 秦川牛ACTC1基因的表达特性分析[J]. 中国畜牧兽医, 2020, 47(8): 2359-2367.
[13]	李京徽, 邢思远, 王希彩, 李庆贺, 赵桂苹, 张永宏, 文杰, 刘冉冉. 鸡肌内脂肪沉积相关候选基因筛选[J]. 中国畜牧兽医, 2020, 47(6): 1828-1836.
[14]	姜徽, 刘攀, 李媛媛, 刘奎国, 黄锡霞, 刘江卫, 胥磊, 杜建文, 桑扎根, 杨雪梅, 赵番番, 王丹. 新疆褐牛乳房炎抗性相关基因的表达及生物信息学分析[J]. 中国畜牧兽医, 2020, 47(1): 90-97.
[15]	张继, 易鸣, 程朝友, 程均华, 付正仙, 杨德文, 吴雪, 邱淦远, 刘若余. 威宁绵羊SOCS7和GFAP基因组织表达研究[J]. 《中国畜牧兽医》, 2019, 46(7): 1899-1906.