从江香猪FoxO 4基因真核表达载体的构建及其在皮下脂肪前体细胞中的表达

doi:10.11843/j.issn.0366-6964.2017.12.006

畜牧兽医学报 ›› 2017, Vol. 48 ›› Issue (12): 2268-2276.doi: 10.11843/j.issn.0366-6964.2017.12.006

从江香猪FoxO 4基因真核表达载体的构建及其在皮下脂肪前体细胞中的表达

孙成娟^1,3, 许厚强^1,2,3*, 段志强^1,3, 陈伟^1,4, 赵佳福^1,4, 周迪^1,4

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:2017-05-25 出版日期:2017-12-23 发布日期:2017-12-23
通讯作者: 许厚强,博士,教授,主要从事细胞分子生物学研究,E-mail:gzdxxhq@163.com
作者简介:孙成娟(1990-),女,山东海阳人,硕士生,主要从事动物遗传育种与种质资源创新研究,E-mail:1254219511@qq.com
基金资助:
国家科技支撑计划（2015BAD03B02-3）；贵州畜禽种质资源保存创新与利用项目（黔科合重大专项字（2013）6008）

Construction of Recombinant Eukaryotic Expression Vector of FoxO 4 Gene from Congjiang Xiang Pig and Its Expression in Subcutaneous Preadipocytes

SUN Cheng-juan^1,3, XU Hou-qiang^1,2,3*, DUAN Zhi-qiang^1,3, CHEN Wei^1,4, ZHAO Jia-fu^1,4, ZHOU Di^1,4

1. Key Laboratory of Animal Genetics, Breeding and Reproduction in the Plateau Mountainous Region of 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:2017-05-25 Online:2017-12-23 Published:2017-12-23

摘要/Abstract

摘要：

本研究旨在克隆从江香猪FoxO 4基因编码区序列，对FoxO 4基因功能进行初步探究。采用qRT-PCR技术对从江香猪（6月龄）不同组织中FoxO 4基因的相对表达量进行检测；PCR扩增FoxO 4基因CDS区，构建真核表达载体pEGFP-N3-FoxO4；利用脂质体法将重组质粒pEGFP-N3-FoxO4瞬时转染从江香猪皮下脂肪前体细胞，通过qRT-PCR方法，检测FoxO 4、PPARγ、LPL、FAS、ACC、ATGL、HSL的表达水平。结果显示：FoxO 4基因在从江香猪脂肪组织中表达量最高；构建的重组真核表达载体pEGFP-N3-FoxO4在从江香猪皮下脂肪前体细胞中成功表达，与对照组相比，LPL、FAS、ACC、ATGL、HSL基因的表达均显著上调。综上表明，FoxO 4对脂质代谢具有一定的调控作用，可能是影响从江香猪猪肉品质的重要候选基因，为进一步了解从江香猪脂肪沉积机制提供理论基础。

Abstract:

The aim of this study was to clone the coding sequence of FoxO 4 gene of Congjiang Xiang pig and preliminarily explore the function of FoxO 4 gene. The relative expression level of FoxO 4 gene was detected by qRT-PCR in different tissues of Congjiang Xiang pig at the age of 6 month.The recombinant plasmid pEGFP-N3-FoxO4 was constructed by inserting the FoxO 4 gene coding sequence into pEGFP-N3, and then transiently transfected into the subcutaneous preadipocytes of Congjiang Xiang pig by liposome-mediated transfection.The expression levels of FoxO 4, PPARγ, LPL, FAS, ACC, ATGL, HSL in transfected cells were detected by qRT-PCR. The results showed that FoxO 4 gene had the highest expression level in adipose tissue. The constructed plasmid pEGFP-N3-FoxO4 was successfully expressed in subcutaneous preadipocytes. The expression level of LPL, FAS, ACC, ATGL and HSL genes were significantly up-regulated in the subcutaneous preadipocytes when compared to the control group.These results revealed that FoxO 4 gene had certain regulation on fat deposition and might be a key candidate gene for the pork quality, which will provide theoretical basis for further studying the mechanism of fat deposition.

中图分类号:

S828.2

孙成娟, 许厚强, 段志强, 陈伟, 赵佳福, 周迪. 从江香猪FoxO 4基因真核表达载体的构建及其在皮下脂肪前体细胞中的表达[J]. 畜牧兽医学报, 2017, 48(12): 2268-2276.

SUN Cheng-juan, XU Hou-qiang, DUAN Zhi-qiang, CHEN Wei, ZHAO Jia-fu, ZHOU Di. Construction of Recombinant Eukaryotic Expression Vector of FoxO 4 Gene from Congjiang Xiang Pig and Its Expression in Subcutaneous Preadipocytes[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2017, 48(12): 2268-2276.

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从江香猪FoxO 4基因真核表达载体的构建及其在皮下脂肪前体细胞中的表达

Construction of Recombinant Eukaryotic Expression Vector of FoxO 4 Gene from Congjiang Xiang Pig and Its Expression in Subcutaneous Preadipocytes

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