Reproductive and Developmental Biology

The Korean Society of Animal Reproduction (한국동물번식학회)
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Development of Low Density Lipoprotein Receptor-Related Protein 5 (LRP5) Gene Targeted Mouse

저밀도 리포단백질 수용체 관련 단백질 5(LRP5) 유전자 적중 생쥐의 개발

  • Park H. Y. (Department of Animal Science, Insti. of Ag. Sci. and Tech., College of Agriculture & Life Science, Chonnam National University) ;
  • Kim C. M. (Department of Animal Science, Insti. of Ag. Sci. and Tech., College of Agriculture & Life Science, Chonnam National University) ;
  • Lee S. M. (Department of Animal Science, Insti. of Ag. Sci. and Tech., College of Agriculture & Life Science, Chonnam National University) ;
  • Jeoung Y. H. (Department of Animal Science, Insti. of Ag. Sci. and Tech., College of Agriculture & Life Science, Chonnam National University) ;
  • Moon S. J. (Department of Animal Science, Insti. of Ag. Sci. and Tech., College of Agriculture & Life Science, Chonnam National University) ;
  • Kang M. J. (Department of Animal Science, Insti. of Ag. Sci. and Tech., College of Agriculture & Life Science, Chonnam National University)
  • 박효영 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 김철민 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 이상미 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 정영희 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 문승주 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부) ;
  • 강만종 (전남대학교 농업생명과학대학, 농업과학기술연구소, 동물자원학부)
  • Published : 2005.03.01
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Abstract

The low density lipoprotein receptor-related protein 5 (LRP5) highly expressed in many tissues, including hepatocytes and pancreatic beta cells, can bind to apolipoprotein E. To evaluate in vivo roles of LRP5, we generated LRP5-deficient mice. LRP5 genomic DNA was isolated from TT2 embryonic stem (ES) cells. Targeting vector was constructed to disrupt an exon 18 of the mouse LRP5 gene and transfected into ES cells. Three homologous recombinants at LRP5 locus were identified from 178 G418-resistant clones. Chimeric males generated by morula aggregation technique were mated to C57BL/6 female mice. After achieving germ-line transmission, LRP5+/- females were crossed with LRP5+/- males to obtain LRP5-deficient mice. One line of mice lacking LRP5 gene was confirmed by Southern blotting. Such knock-out mice may serve as an effective animal model to study in vivo function of LRP5 gene.

저밀도 리포단백질 수용체 관련 단백질 5(LRP5)는 간과 췌장을 포함하여 많은 조직에서 발현하며 아포리포단백질 E와 결합한다. 이와 같은 LRP5 유전자의 체내 기능을 규명하기 위하여 LRP5 유전자가 결손된 생쥐를 개발하였다. 먼지 LRP5 genomic DNA는 TT2 ES 세포로부터 분리하였으며 LRP5 유전자의 엑손 18에 neo 유전자를 삽입한 vector를 구축하고 TT2 ES 세포에 도입하였다. 178개의 G418 내성을 보인 세포 중 상동유전자 재조합에 의하여 targeting vector가 LRP5 유전자 위치에 삽입된 clone은 3개였다. 키메라 생쥐는 상실배기 수정을 ES 세포와 응집시켜 생산하였으며 생산된 키메라 생쥐는 C57BL/6 생쥐와 교미를 유도하여 heterozygous를 얻었다. 또한 이들 heterozygous간의 교배에 의하여 LRP5 유전자 결손 생쥐를 생산하였다. 이러한 생쥐는 LRP5 유전자의 체내 기능연구에 있어서 모델로 이용될 것으로 생각된다.

Keywords

References

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