Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
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A New Bicistronic Fragmentation Vector for Manipulation and Analysis of Functional Yeast Artificial Chromosomes (YACs)

Yeast Artificial Chromosome의 효율적인 조작과 분석을 위한 새로운 Bicistronic Fragmentation Vector의 개발에 관한 연구

  • 임향숙 (가톨릭대 의과학연구원 분자유전학연구소) ;
  • 최주연 (가톨릭대 의과학연구원 분자유전학연구소) ;
  • 김인경 (가톨릭대 의과학연구원 분자유전학연구소) ;
  • 강성만 (고려대 생명공학원) ;
  • 성영모 (고려대 생명공학원)
  • Published : 1999.03.01
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Abstract

Fragmentation vectors are used to analyze function and genomic structure of a gene of interest by creating deletion derivatives of large fragments of genomic DNA cloned as yeast artificial chromosomes (YACs). Herein, we developed a new hicistronic fragmentation vector that contains internal ribosomal entry sile (IRES) of encephalomyocarditis vin~s (EMCV) and $\beta$-galactosidase as a reporter gene. This vector system provides a novcl loo1 to analyze expression patterns of a gene of interest due to simultaneous expression of a target gene as well as $\beta$-galactosidase driven from a single message. In addition, the bicistronic fragmentation vector contains four rare-cutting restriction enzyme sites in the polycloning sites which can be used to conveniently insert any kinds of genes and therefore facilitates targeting DNA scgments into YAC by means of homologous recombination. This approach establishes a paradigm for manipulation of mammalian DNA segments and characterization of expression and regulatory regions of mammalian gene cloned as YAC.

본 연구에서는 Yeast Artificial Chromosome을 효율적으로 조작하고 유유전자의 기능을 보다 더 용이하게 분석하기 위해 EMCV 바이러스의 IRES 염기서열과 $\beta$-galactosidase를 포함하는 새로운 bicistronic fragmentation vector를 제조하였다. 이 벡터의 폴리클로닝 site에 네 개의희귀한 제한효소 부위를 도입하여 DNA를 용이하게 클로닝할 수 있게 만들었다. 그러므로 원하는 어떤 YAC도 효모세포에서 쉽게 상동 재조함에 의해 절편할 수 있는 장점이 있다. 이 bicistronic fragmentation vector 시스템을 이용하면하나의 메시지로부터 연구하고자 하는 유전자와 $\beta$-galactosidase를 동시에 한 세포에서 발현할 수 있기 때문에 유전자의 발현양상을 쉽게 분석할 수 있는 새로운 도구로 이용할 수 있다.

Keywords

References

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