广西巴马小型猪PGC-1α基因克隆与组织表达分析
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广西自然科学基金项目(2013GXNSFAA019187);国家自然科学基金项目(81360135).


Cloning and analysis of tissue expression of PGC-1α gene in Guangxi Bama mini-pigs
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    摘要:

    目的 克隆广西巴马小型猪PGC-1α基因编码区(CDS)序列,利用RT-PCR和QRT-PCR方法分析PGC-1α mRNA组织表达情况.方法 本实验以广西巴马小型猪背最长肌cDNA为模版,PCR扩增PGC-1α基因CDS序列,将其连接至pEASY-T5载体,转染细菌、验证和序列测定;通过RT-PCR半定量和QRT-PCR实时荧光定量检测PGC-1α基因在小型猪多个组织中的表达情况.结果 克隆获得广西巴马小型猪PGC-1α基因CDS序列,全长2391 bp,编码796个氨基酸,与参考序列的同源性为99.9%,两处碱基发生同义突变,分别是C-A1105和G-A1524;PGC-1α基因在广西巴马小型猪心脏和肾脏中的表达丰度最高,其次是肝脏、皮下脂肪和背最长肌,而在胰腺中未检测到其表达.结论 成功克隆了广西巴马小型猪PGC-1α基因编码区序列并进行了多种组织表达分析,为后续研究PGC-1α在小型猪2型糖尿病发生过程中作用途径打下基础.

    Abstract:

    Objective To clone the coding sequence of Guangxi Bama mini-pig PGC-1α gene, and to analyze the expression of PGC-1α gene in various tissues of mini-pigs using RT-PCR and QRT-PCR techniques. Methods The PGC-1α gene coding sequence (CDS) was amplified by PCR from the cDNA of longissimus muscle of Guangxi Bama mini-pig. The PCR products were inserted into pEASY-T5 vector, transfected E. coli, identified and sequenced. The PGC-1α gene expression in different tissues of the Bama mini-pigs was detected by RT-PCR and QRT-PCR assays. Results The PGC-1α gene CDS of Guangxi Bama mini-pig was cloned. It was 2391 bp in length. It had 99.9% homology with the reference sequence, and had two synonymous mutations that were C-A1105 and G-A1524. The expression level of PGC-1α gene was higher in the heart and kidney, followed by liver, subcutaneous fat and longissimus muscle, but the expression was not detected in pancreas of Guangxi Bama mini-pig. Conclusions We have successfully cloned the PGC-1α gene of Guangxi Bama mini-pig, and detected this gene expression in six tissues. The results of this study will provide a basis for studying the effect of PGC-1α on type 2 diabetes mellitus (T2DM) in Bama mini-pigs.

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严雪瑜,敖秋桅,严小东,蒋钦杨,郭亚芬,兰干球.广西巴马小型猪PGC-1α基因克隆与组织表达分析[J].中国实验动物学报,2014,22(5):27~31.

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  • 收稿日期:2014-04-25
  • 在线发布日期: 2014-10-30
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