JARB Journal of Animal Reproduction and Biotehnology

OPEN ACCESS pISSN: 2671-4639
eISSN: 2671-4663

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Reproductive & Developmental Biology 2003; 27(1): 15-23

Published online March 30, 2003

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

Mouse Mammary Epithelial Cell에서 Retrovirus Vector를 이용한 Human Lactadherin 유전자의 유도적 발현

권모선,구본철,정병현,염행철,박창식,김태완

대구가톨릭대학교 의과대학 생리학교실,건국대학교 수의과대학,호서대학교 생명과학전공,충남대학교 동물자원과학부, 형질전환복제돼지 연구센터

Abstract

Lactadherin (formerly known as BA46), a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and breast carcinomas and may prevent symptomatic rotavirus infections. In this study, under the control of tissue specific and hormonal inducible mouse whey acidic protein (WAP) promote., the expression pattern of lactadherin (Ltd) in lactogenic hormone-dependent mouse mammary epithelial cell line HC11 were tested. pLNWLtd construct containing 2.4 kilobases of the WAP promoter and 1.5 kilobases of human lactadherin gene was stably transfered into HC11 cells using retroviral vector system. Integration and expression level of the transgene was estimated using PCR and RT-PCR, respectively. Prominent induction of Ltd gene under the WAP promoter was accomplished in the presence of insulin, hydrocortisone and prolactin. Compared to the control (cells cultured with insulin alone), however we observed that the WAP promoter was leaky. These data indicate that luther studies are needed in finding an appropriate promoter other than WAP promoter because of its leakiness.

Keywords: Human lactadherin gene, Whey acidic protein promoter, Lactogenic hormones, RT-PCR

Article

Reproductive & Developmental Biology 2003; 27(1): 15-23

Published online March 30, 2003

Copyright © The Korean Society of Animal Reproduction and Biotechnology.

Mouse Mammary Epithelial Cell에서 Retrovirus Vector를 이용한 Human Lactadherin 유전자의 유도적 발현

권모선,구본철,정병현,염행철,박창식,김태완

대구가톨릭대학교 의과대학 생리학교실,건국대학교 수의과대학,호서대학교 생명과학전공,충남대학교 동물자원과학부, 형질전환복제돼지 연구센터

Abstract

Lactadherin (formerly known as BA46), a major glycoprotein of the human milk fat globule membrane, is abundant in human breast milk and breast carcinomas and may prevent symptomatic rotavirus infections. In this study, under the control of tissue specific and hormonal inducible mouse whey acidic protein (WAP) promote., the expression pattern of lactadherin (Ltd) in lactogenic hormone-dependent mouse mammary epithelial cell line HC11 were tested. pLNWLtd construct containing 2.4 kilobases of the WAP promoter and 1.5 kilobases of human lactadherin gene was stably transfered into HC11 cells using retroviral vector system. Integration and expression level of the transgene was estimated using PCR and RT-PCR, respectively. Prominent induction of Ltd gene under the WAP promoter was accomplished in the presence of insulin, hydrocortisone and prolactin. Compared to the control (cells cultured with insulin alone), however we observed that the WAP promoter was leaky. These data indicate that luther studies are needed in finding an appropriate promoter other than WAP promoter because of its leakiness.

Keywords: Human lactadherin gene, Whey acidic protein promoter, Lactogenic hormones, RT-PCR