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Inducible Expression of the Lactadherin Gene with a Reverse Tetracycline-Regulated Retroviral Vector System  

이용석 (대구가톨릭대학교 의과대학 병리학교실)
오훈규 (계명대학교 의과대학 병리학교실)
권모선 (대구가톨릭대학교 의과대학 생리학교실)
박창식 (충남대학교 동물자원과학부, 충남대학교 형질전환복제돼지 연구센터)
김태완 (대구가톨릭대학교 의과대학 생리학교실, 충남대학교 형질전환복제돼지 연구센터)
박재복 (대구가톨릭대학교 의과대학 병리학교실)
Publication Information
Korean Journal of Animal Reproduction / v.27, no.3, 2003 , pp. 259-268 More about this Journal
Abstract
Lactadherin (formerly known as BA46), a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and breast carcinoma cells and is known to prevent symptomatic rotavirus infections. In this study, we tried to transfer the human lactadherin gene to the Chinese Hamster Ovary (CHO) cells using retrovirus vector system and tested inducible expression of the gene under the tetracycline-controllable promoter. At first, tetracycline-mediated inducibility was tested using E.coli LacZ marker gene. NIH3T3 cells co-infected with RevTet-On and RevTRE-LacZ retrovirus vectors showed that the cells responded to doxycycline (a derivative of tetracycline) in a dose-dependent manner, and prominent induction of the lacZ gene expression was observed from 1 $\mu\textrm{g}$/ml of doxycycline concentration. Based on the results of the pilot experiment, inductional expression of the human lactadherin gene was conducted using RevTet-On and RevTRE-Ltd retrovirus vectors. Analysis with the RT-PCR demonstrated successful inductional expression of the lactadherin gene in the Chinese Hamster Ovary (CHO) cells. Considering that constitutive overexpression of the exogenous genes in the target cells causes serious physiological imbalance, the results obtained in this study will be very useful especially in the studies of gene therapy and transgenic animal production.
Keywords
Ltd gene; Inducible expression; Doxycycline; CHO cell; RT-PCR;
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