Journal of Life Science (생명과학회지)

Korean Society of Life Science (한국생명과학회)
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Effect of random Shine-Dalarno sequence on the expression of Bovine Growth Hormone Gene in Escherichia coli

대장균에서 무작위 샤인-달가노 서열이 소성장호르몬 유전자 발현에 미치는 영향

  • 나경수 (대구공업대학 식품영양과) ;
  • 나경수 (대구공업대학 식품영양과) ;
  • 백형석 (부산대학교 미생물학과) ;
  • 이용세 (대구대학교 자연자원대학 생명자원학부)
  • Published : 2000.08.01
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Abstract

In order to search for the effects of Shine-Dalgarno (SD) sequence and nucleotide sequence of spacer region (SD-ATG) on bGH expression, oligonucleotides containing random SD sequences and a spacer region were chemically synthesized. The distance between SD region and initiation codon (ATG) was fixed to 9 nucleotides in length. The expression vectors have been constructed using pT7-1 vector containing a T7 promoter. Positive clones were screened with colony hybridization and named pT7A or pT7B plasmid series. The selected clones were confirmed by DNA sequencing and finally, 19 clones having various SD combinations were obtained. When bovine growth hormone was induced by IPTG in E. coli BL21(DE3), all cells harboring these plasmids produced a detectable level of bGH in western blot analysis. However, various SD sequences did not affect on bGH expression, indicating that the sequences of SD and the spacer region did not sufficiently destabilize mRNA secondary structure of bGH gene. Therefore, these results indicate that the disruption of mRNA secondary structure might be a major factor for regulating bGH expression in the translational initiation process.

Keywords

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