원문정보
Production of a Transgenic Pig Overexpressing Phosphoprotein Enriched in Astrocytes 15 (PEA 15)
초록
영어
The overexpression of Phosphoprotein Enriched in Astrocytes (PEA15) gene is commonly found in human diabetic patients. The overexpression of this gene in skeletal muscle and fat tissues have been reported to cause insulin resistance, thereby impairing insulin stimulated glucose uptake. We introduced a gene of mouse PEA15 (mPEA15) and enhanced green fluorescent protein (EGFP) into fertilized one cell pig zygotes using microinjection, and produced a piglet that showed overexpression of mPEA15 in the muscle tissues and expression of EGFP in the ear tissues and hooves. RT-PCR RFLP, southern blot and FISH analysis showed that the tissues carried the transgene. Real-time RT-PCR and western blots demonstrated that PEA15 gene was overexpressed in the various tissues and muscle tissues, respectively. These facts suggest that expression vector system is normally expressed in the trnasgenic (TG) pigs. To use as animal diseases model for type 2 diabetes, further study is necessary to confirm whether diabetes occur in these TG pigs, especially insulin resistance.
목차
서론
재료 및 방법
미세 주입용 유전자 준비
Micropipets 제작 및 유전자 미세 주입
외과적 방법에 의한 수정란 채란 및 이식
RT-PCR RFLP 및 Real-time RT-PCR
Southern Blot Hybridization
Western Blotting
Fluorescence In situ Hybridization (FISH)
결과 및 고찰
mPEA15의 과발현 및 EGFP 발현의 확인
인용문헌