원문정보
In Vitro Assay of Mammary Gland Tissue Specific hEPO Gene Expression
초록
영어
Effectiveness of transgene transfer into genome is crucially concerned in mass production of the bio-pharmaceuticals using genetically modified transgenic animals as a bioreactor. Recently, the mammary gland has been considered as a potential bioreactor for the mass production of the bio-pharmaceuticals, which appears to be capable of appropriate post-translational modifications of recombinant proteins. The mammary gland tissue specific vector system may be helpful in solving serious physiological disturbance problems which have been a major obstacle in successful production of transgenic animals. In this study, to minimize physiological disturbance caused by constitutive over-expression of the exogenous gene, we constructed new retrovirus vector system designed for mammary gland-specific expression of the hEPO gene. Using piggyBac vector system, we designed to express hEPO gene under the control of mammary gland tissue specific and lactogenic hormonal inducible goat β-casein or mouse Whey Acidic Protein (mWAP) promoter. Inducible expression of the hEPO gene was confirmed using RT-PCR and ELISA in the mouse mammary gland cells treated with lactogenic hormone. We expect the vector system may optimize production efficiency of transgenic animal and reduce the risk of global expression of transgene.
목차
서론
실험 방법
hEPO 유전자가 유선조직 특이적으로 발현되는 Vector의 구축
pPiggy-βCaseinp-hEPO-Puro가 도입된 세포주의 구축 및 유선조직 특이적인 hEPO 유전자 발현 유도
RT-PCR 방법을 이용한 유선조직 특이적인 hEPO 유전자 발현의 In Vitro 검정
ELISA 방법을 이용한 유선조직 특이적인 hEPO 유전자 발현의 In Vitro 검정
pPiggy-mWAPp-hEPO-EF1-Puro가 도입된 세포주의 구축및 유선조직 특이적인 hEPO 유전자 발현 검정
세포에서 생산된 재조합 hEPO의 생물학적 활성 검정
결과
pPiggy-βCaseinp-hEPO-EF1-Puro Vector를 이용한 유선조직 특이적인 hEPO 유전자 발현의 In Vitro 검정
pPiggy-mWAPp-hEPO-EF1-Puro Vector를 이용한 유선조직 특이적인 hEPO 유전자 발현의 In Vitro 검정
고찰
REFERENCES