Analysis of Tissue-Specific Expression of Human Type II Collagen cDNA Driven by Different Sizes of the Upstream Region of the .BETA.-Casein Promoter

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  • NARUSE Kenji
    Research Center for Transgenic and Cloned Pigs, Chungnam National University
  • YOO Seung Kwon
    Division of Biotechnology and Genetic Engineering, Korea University
  • KIM Sun Myoung
    Department of Animal Science and Technology, Seoul National University
  • CHOI Yun Jaie
    Department of Animal Science and Technology, Seoul National University
  • LEE Hong Mie
    Department of Food Science and Nutrition, Daejin University
  • JIN Dong Il
    Research Center for Transgenic and Cloned Pigs, Chungnam National University

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Other Title
  • Analysis of Tissue-Specific Expression of Human Type 2 Collagen cDNA Driven by Different Sizes of the Upstream Region of the β-Casein Promoter
  • Analysis of Tissue Specific Expression of Human Type 2 Collagen cDNA Driven by Different Sizes of the Upstream Region of the ベータ Casein Promoter
  • Analysis of Tissue-Specific Expression of Human Type II Collagen cDNA Driven by Different Sizes of the Upstream Region of the β-Casein Promoter

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Abstract

To investigate the ability of 1.8 kb or 3.1 kb bovine β-casein promoter sequences for the expression regulation of transgene in vivo, transgenic mice were produced with human type II collagen gene fused to 1.8 kb and 3.1 kb of bovine β-casein promoter by DNA microinjection. Five and three transgenic founder mice were produced using transgene constructs with 1.8 kb and 3.1 kb of bovine β-casein promoters respectively. Founder mice were outbred with the wild type to produce F1 and F2 progenies. Total RNAs were extracted from four tissues (mammary gland, liver, kidney, and muscle) of female F1 transgenic mice of each transgenic line following parturition. RT-PCR and Northern blot analysis revealed that the expression level of transgene was variable among the transgenic lines, but transgenic mice containing 1.8 kb of promoter sequences exhibited more leaky expression of transgene in other tissues compared to those with 3.1 kb promoter. Moreover, Western blot analysis of transgenic mouse milk showed that human type II collagen proteins secreted into the milk of lactating transgenic mice contained 1.8 kb and 3.1 kb of bovine β-casein promoter. These results suggest that promoter sequences of 3.1 kb bovine β-casein gene can be used for induction of mammary gland-specific expression of transgenes in transgenic animals.

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