Contact Us Add to Favorite
 
NianQi Search Adv Search
33
研究资源 Current Issue | Archive | Adv Search |
Construction and Evaluation of Porcine Adenosine Monophosphate Activated Protein Kinase α Gene(AMPKα) siRNA Expression Vectors
Download: PDF (545 KB)   HTML (1 KB) 
Export: BibTeX | EndNote (RIS)      
Abstract  The adenosine monophosphate activated protein kinase(AMPK)is a key regulator of catabolic and anabolic processes in energy metabolism. This study was conducted to construct and select the effective siRNA interference vector for porcine AMPKα gene. Two pairs of porcine AMPK α-specific double-strand siRNA(dsRNA) primers were designed and inserted into pSilencer 4.1-CMV neo Vector after annealing. The vectors were then transfected into porcine intramuscular preadipocytes by lipofection 2000, AMPKα and lipolitic enzyme genes HSL(hormone sensitive lipase), ATGL(adipose triglyceride lipase) and CPT1(carnitine palmitoyl transferase) mRNA expressions in these cells were detected by Real-time PCR. The results showed that the AMPKα-specific siRNA vectors RA1 and RA2 were successfully constructed and transfected into porcine intramuscular preadipocytes. The efficiency of RA2 was significantly higher than that of RA1. The mRNA expressions of AMPKα and the lipolitic enzyme genes HSL, ATGL and CPT1 decreased significantly after treatment of porcine intramuscular preadipocytes by vector RA2(P<0.05). It indicated that decomposition of fat and fatty acid oxidation rate decreased in porcine intramuscular preadipocytes when AMPKα mRNA expressions was inhibited. These data will provide insight in the function of AMPK in fat deposition and manipulating gene expression of AMPK in regulating fat metabolism and meat quality in pigs.
Key wordsAdenosine monophosphate activated protein kinase(AMPK)      Construction of expression vector      Porcine      Interference effect     
Received: 06 May 2013     
Service
E-mail this article
Add to my bookshelf
Add to citation manager
E-mail Alert
RSS
Articles by authors
NULL
Cite this article:   
NULL. Construction and Evaluation of Porcine Adenosine Monophosphate Activated Protein Kinase α Gene(AMPKα) siRNA Expression Vectors[J]. , 2013, 21(9): 1119-1124.
URL:  
http://journal05.magtech.org.cn/Jwk_ny/EN/     OR     http://journal05.magtech.org.cn/Jwk_ny/EN/Y2013/V21/I9/1119
Copyright © Editorial Board of 农业生物技术学报
Supported by:Beijing Magtech