InVitro에서 RetrovirusVectorSystem을 이용한 재조합 hFSH 유전자의 발현

서지정보

ㆍ발행기관 : 한국동물번식학회 ㆍ수록지정보 : Reproductive & developmental biology / 35권 / 1호
ㆍ저자명 : 민경헌, 권모선, 김태완, 구본철

목차

ABSTRACT 서론 실험 방법 Retrovirus의 구축 Virus의 생산 및 표적 세포로의 감염 재조합 hFSH 유전자 및 hFSHR 유전자 발현 확인 ELISA를 이용한 재조합 hFSH의 정량 재조합 hFSH의 생물학적 활성 측정 결과 Retrovirus Vector System을 이용한 재조합 hFSH 유전자발현 확인 Retrovirus Vector System을 이용한 hFSHR 유전자 발현확인 재조합 hFSH의 생물학적 활성 측정 고찰 인용문헌

영어 초록

hFSH is a glycoprotein secreted from anterior pituitary and consists of α and β subunits. Because of its major biological functions including sperm formation in the male and for follicular growth, FSH is used to cure woman's sterility. In this study we tried to produce recombinant hFSH in vitro using a retrovirus expression vector. Two major components of the vector we constructed are: (ⅰ) a DNA fragment containing α and β genes fused by a DNA sequence coding carboxyl terminal peptide (CTP) of human chorionic gonadotropin, (ⅱ) a DNA fragment corresponding woodchuck hepatitis virus posttranscriptional regulatory element (WPRE). Evaluation of expression profile of the recombinant FSH using reverse transcription PCR and enzyme-linked immunosorbent assay (ELISA). Among three cell lines tested, HeLa cells were the best for hFSH expression (5,395 mIU/ml), then followed by chicken embryonic fibroblast (CEF) cells and Chinese hamster ovary (CHO) cells in the order of hFSH production. In addition to the amount, the FSH produced from HeLa cells was highest in terms of biological activity which was determined by measuring cAMP.

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