Analysis of tissue-specific expression of human type II collagen cDNA driven by different sizes of the upstream region of the β-casein promoter

Kenji Naruse, Kwon Yoo Seung, Myoung Kim Sun, Jaie Choi Yun, Mie Lee Hong, Dong Il Jin

Research output: Contribution to journalArticle

11 Citations (Scopus)

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 micro-injection. 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.

Original languageEnglish
Pages (from-to)93-98
Number of pages6
JournalBioscience, Biotechnology and Biochemistry
Volume70
Issue number1
DOIs
Publication statusPublished - 2006

Keywords

  • Bovine β-casein promoter
  • Gene expression
  • Human type II collagen gene
  • Milk
  • Transgenic mice

ASJC Scopus subject areas

  • Biotechnology
  • Analytical Chemistry
  • Biochemistry
  • Applied Microbiology and Biotechnology
  • Molecular Biology
  • Organic Chemistry

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