伊犁马PFKM基因表达及酶活性分析

doi:10.6048/j.issn.1001-4330.2022.11.020

新疆农业科学 ›› 2022, Vol. 59 ›› Issue (11): 2758-2764.DOI: 10.6048/j.issn.1001-4330.2022.11.020

• 马运动生理与健康养殖 • 上一篇下一篇

伊犁马PFKM基因表达及酶活性分析

王洁¹, 孟军¹^,², 曾亚琦¹^,², 王建文¹^,², 吐尔逊江·吾木尔艾力³, 王川坤¹, 袁鑫鑫¹, 王彤亮¹, 姚新奎¹^,²()

1.新疆农业大学动物科学学院/新疆农业大学马产业研究院,乌鲁木齐 830052
2.新疆马繁育与运动生理重点实验室,乌鲁木齐 830052
3.新疆伊犁哈萨克自治州畜牧总站,新疆伊宁 835000

收稿日期:2022-01-11 出版日期:2022-11-20 发布日期:2022-12-28
通信作者: 姚新奎
作者简介:王洁(1996-),女,新疆伊犁人,硕士研究生,研究方向为动物生产学,(E-mail)1547628788@qq.com
基金资助:
新疆维吾尔自治区天池博士计划项目;新疆维吾尔自治区重大科技专项(2022A02007-1);自治区创新环境(人才、基地)建设专项(PT2220);新疆伊犁马产业集群项目(2022xjmlt-z-01)

The mRNA Expression and Activity of PFKM in Different Tissues of Yili Horses

WANG Jie¹, MENG Jun¹^,², ZENG Yaqi¹^,², WANG Jianwen¹^,², Tuerxunjiang Wumuraili³, WANG Chuankun¹, YUAN Xinxin¹, WANG Tongliang¹, YAO Xinkui¹^,²()

1. College of Animal Science,Xinjiang Agricultural University/Research Institute of Horse Industry,Xinjiang Agricultural University,Urumqi 830052,China
2. Xinjiang Key Laboratory of Equine Breeding and Exercise Physiology,Urumqi 830052,China
3. Xinjiang Yili Kazakh Autonomous Prefecture Animal Husbandry Station, Yining Xinjiang 835000,China

Received:2022-01-11 Online:2022-11-20 Published:2022-12-28
Correspondence author: YAO Xinkui
Supported by:
Dr. Tianchi Project of Xinjiang Uygur Autonomous Region;Major science and technology projects of Xinjiang Uygur Autonomous Region(2022A02007-1);The Innovation Environment (Talent, Base) Construction Project of Xinjiang Uygur Autonomous Region(PT2220);Xinjiang Yili Horse Industry Cluster Project(2022xjmlt-z-01)

摘要/Abstract

摘要：

【目的】测定PFKM基因在伊犁马不同组织部位中的差异表达,为该基因在伊犁马的糖酵解过程中功能提供一定基础。【方法】以伊犁马为试验动物,分别提取心脏、肝脏、夹肌、斜方肌、背阔肌、臀中肌、半腱肌以及腹外斜肌8种组织的总RNA,以伊犁马不同组织的RNA为模板,逆转录合成cDNA,GAPDH为内参基因,运用荧光定量PCR的方法检测PFKM基因mRNA在伊犁马不同组织间的相对表达量。运用酶联免疫法(Elisa)检测伊犁马不同组织中糖酵解相关酶的含量。【结果】伊犁马PFKM基因在以上8种组织部位中都有表达,在心脏中相对表达量最高,肝脏中相对表达量最低,心脏中PFKM基因mRNA相对表达量极显著高于马匹其他部位(P<0.01)。马匹心脏组织中6-磷酸果糖激酶的活性显著高于其他部位。【结论】在糖酵解途径中,PFKM基因6-磷酸果糖激酶对心脏组织的糖酵解有重要的影响。

关键词: 伊犁马; PFKM; 糖酵解; 6-磷酸果糖激酶; 表达

Abstract:

【Objective】 To determine the differential expression of PFKM gene in different tissue sites in Yili horses in the hope of providing some research basis to clarify the function of this gene during glycolysis in the horses. 【Methods】 Total RNA was extracted from the heart, liver, clip, trapezius, latissimus dorsi, gluteus medius, semitendinosus and external oblique of the abdomen of the Yili Horse, the relative expression of PFKM mRNA in different tissues was detected by real-time fluorescent quantitative PCR. And the cDNA was synthesized by reverse transcription using RNA from different tissues and GAPDH as internal reference gene. Enzyme-linked immunosorbent assay (Elisa) was used to determine the content of related enzymes in tissues. 【Results】 PFKM gene was expressed in all of the above 8 tissues, with the highest expression level in heart tissues and the lowest in liver tissues, the expression of PFKM gene mRNA in heart was significantly higher than that in other tissues (P< 0.01). Analysis of 6-phosphofructokinase activity in eight tissues of Yili Horses showed that 6-phosphofructokinase activity in heart tissue was significantly higher than that in other tissues, suggesting that in glycolytic process. 【Conclusion】 PFKM gene is involved in the regulation of glycolysis through the action of 6- PFK in the heart tissue, the results of which will lay a foundation for analyzing the effect of PFKM gene in glycolytic process.

Key words: Yili Horse; PFKM; glycolysis; 6-PFK; expression

中图分类号:

S821.9

王洁, 孟军, 曾亚琦, 王建文, 吐尔逊江·吾木尔艾力, 王川坤, 袁鑫鑫, 王彤亮, 姚新奎. 伊犁马PFKM基因表达及酶活性分析[J]. 新疆农业科学, 2022, 59(11): 2758-2764.

WANG Jie, MENG Jun, ZENG Yaqi, WANG Jianwen, Tuerxunjiang Wumuraili, WANG Chuankun, YUAN Xinxin, WANG Tongliang, YAO Xinkui. The mRNA Expression and Activity of PFKM in Different Tissues of Yili Horses[J]. Xinjiang Agricultural Sciences, 2022, 59(11): 2758-2764.

图/表 6

表1 引物信息

Table 1 PRIMER INFORMATION

基因 Gene	引物序列 Sequences of primers	退火温度 Annealing temperature(℃)	产物长度 Length
PFKM-F2	ACTTGGGACTAAAAGGACGCTACC	60	51
PFKM-R2	GCACTGATCTGCTCGAAGCTC	60	51
GAPDH-F	GGTGAAGGTCGGAGTAAACG	60	87
GAPDH-R	AATGAAGGGGTCATTGATGG	60	87

图1 总RNA提取结果注:1:夹肌;2:斜方肌;3:背阔肌;4:臀中肌;5:半腱肌;6:肝脏;7:心脏;8:腹外斜肌

Fig.1 Total RNA extraction results Note:1.Clamping; 2.Trapezius; 3.Latissimus Dorsi; 4.Gluteals Medius; 5.Semitendinosus; 6.Liver; 7.Cardiac; 8.Obliques

图2 PFKM基因qRT-PCR 溶解曲线和扩增曲线

Fig.2 qRT-PCR dissolution curve and amplification curve of PFKM gene

图3 GAPDH基因qRT-PCR溶解曲线和扩增曲线

Fig.3 qRT-PCR dissolution curve and amplification curve of GAPDH gene

图4 伊犁马PFKM基因mRNA在不同组织中的相对表达量

Fig.4 Relative expression of PFKM gene mRNA in different tissues of Yili horse

表2 6-PFK在不同组织的含量

Table 2 6-PFK content in different tissues

项目 Project	夹肌 Clamping	斜方肌 Trapezius	背阔肌 Latissimus Dorsi	臀中肌 Gluteals Medius	半腱肌 Semiten dinosus	肝脏 Liver	心脏 Cardiac	腹外斜肌 Obliques
6-PFK (mg/L)	0.263± 0.073^CDde	0.266± 0.088^CDde	0.542± 0.127^Bb	0.336± 0.096^Cc	0.317 ±0.492^CDcd	0.262± 0.076^CDde	1.049± 0.245^Aa	0.253± 0.071^De

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伊犁马PFKM基因表达及酶活性分析

The mRNA Expression and Activity of PFKM in Different Tissues of Yili Horses

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