猪THRSP基因5′侧翼区序列转录调控活性的鉴定

畜牧兽医学报 ›› 2011, Vol. 42 ›› Issue (3): 329-334.doi:

猪THRSP基因5′侧翼区序列转录调控活性的鉴定

陈华^1，2，陈宏权^1，2* ，周倩倩¹，张翼鹏¹，魏汉卿¹，李春苗^1，2，章孝荣^1，2，彭英林³

1. 安徽农业大学动物科技学院，合肥 230036；2. 安徽地方畜禽遗传资源保护与生物育种省级实验室，合肥 230036；3. 湖南省畜牧兽医研究所，长沙 410131

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-03-25 发布日期:2011-03-25
通讯作者: 陈宏权

Identification of Transcription Regulation Activity in 5′Flanking Regionof Pig THRSP Gene

CHEN Hua^1，2, CHEN Hong-quan^1，2*, ZHOU Qian-qian¹, ZHANG Yi-peng¹, WEI Han-qing¹, LI Chun-miao^1，2 , ZHANG Xiao-rong^1，2, PENG Ying-lin³

1. College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036,China; 2. Key Laboratory of Anhui Local Livestock and Poultry Genetic Resources Conservation and Biobreeding of Anhui Province, Hefei 230036, China; 3. Hunan Institute of Animal Science and Veterinary Medicine, Changsha 410131, China

Received:1900-01-01 Revised:1900-01-01 Online:2011-03-25 Published:2011-03-25

摘要/Abstract

摘要： 本研究旨在对猪THRSP基因5′调控区TP526序列进行转录调控活性的鉴定，以鉴别THRSP基因调控区的功能。提取猪耳组织基因组，利用PCR技术克隆THRSP基因5′调控区TP526序列，将其插入荧光素酶报告载体中（pGL3-Basic），构建猪THRSP基因5′调控区TP526序列调控的荧光素酶报告载体（pGL3-TP526 /promoter），经PCR鉴定后，转染293T细胞，利用双报告基因检测TP526序列的启动子活性。结果显示，pGL3-TP526 /promoter调控的表达载体，其萤火虫与海肾荧光素酶荧光强度之比（1.816 2±0.253 3）显著高于pGL3-Basic（0.126 7±0.020 3），呈高效表达（P<0.01）。THRSP基因5′调控区TP526序列具有启动子调控活性。

Abstract: The aims of this study were to identify TP526promoter activity of pigs THRSP gene and to understand functions of 5′ regulation region of THRSP gene. The DNA samples were extracted from pig ear tissue. The TP526promoter of thyroid hormon einducible nuclear protein Spot14 (THRSP) gene was amplified using PCR technology, inserted into the luciferase reporter vector (pGL3-basic) to construct pGL3-TP526/promoter. 293T cells were cotransfected with a plasmid containing the pGL3-TP526/promoter and pGL3-basic, respectively. The expression of luciferase gene in 293T cells was tested using the Dual-Luciferase Reporter Assay kit. The results showed that the fluorescence intensity ratio of firefly to sea pansy luciferase with （1.816 2±0.253 3） of pGL3-TP526/promoter was significant higher than that with （0.126 7±0.020 3） of pGL3-basic (P<0.01). It implied a significant promoter activity of TP526.

陈华;陈宏权;;周倩倩;张翼鹏;魏汉卿;李春苗;章孝荣;彭英林. 猪THRSP基因5′侧翼区序列转录调控活性的鉴定[J]. 畜牧兽医学报, 2011, 42(3): 329-334.

CHEN Hua;CHEN Hong-quan;ZHOU Qian-qian;ZHANG Yi-peng;WEI Han-qing;LI Chun-miao;;ZHANG Xiao-rong;PENG Ying-lin . Identification of Transcription Regulation Activity in 5′Flanking Regionof Pig THRSP Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2011, 42(3): 329-334.