• 검색어에 아래의 연산자를 사용하시면 더 정확한 검색결과를 얻을 수 있습니다.
  • 검색연산자
검색연산자 기능 검색시 예
() 우선순위가 가장 높은 연산자 예1) (나노 (기계 | machine))
공백 두 개의 검색어(식)을 모두 포함하고 있는 문서 검색 예1) (나노 기계)
예2) 나노 장영실
| 두 개의 검색어(식) 중 하나 이상 포함하고 있는 문서 검색 예1) (줄기세포 | 면역)
예2) 줄기세포 | 장영실
! NOT 이후에 있는 검색어가 포함된 문서는 제외 예1) (황금 !백금)
예2) !image
* 검색어의 *란에 0개 이상의 임의의 문자가 포함된 문서 검색 예) semi*
"" 따옴표 내의 구문과 완전히 일치하는 문서만 검색 예) "Transform and Quantization"
쳇봇 이모티콘
ScienceON 챗봇입니다.
궁금한 것은 저에게 물어봐주세요.

논문 상세정보

Overexpression and Purification of Reverse Transcriptase of Retron EC83 by Changing the Downstream Sequence of the Initiation Codon


Retron is a prokaryotic genetic element, producing a short single-stranded DNA covalently linked to RNA (msDNA-RNA) by a reverse transcriptase (RT). In retron EC83, msDNA is further processed at between the 4th and the $5^{th}$ nucleotides, leaving a 79 nucleotide-long single-stranded DNA as a final product. To investigate this site-specific cleavage in msDNA synthesis, we purified the RT protein of retron EC83. Initially, RT ORF was cloned under the tac promoter, but the expression was very poor largely because of poor translation. In order to facilitate translation, the nucleotide sequence for the first nine amino acids was randomized with synonymous codons. This change of downstream sequence of translational initiation codon greatly affected the efficiency of translation. We could isolate clones which greatly increased RT production, and their sequences were compared to those of the low producers. The overproduced protein was purified and was shown to have RT activity.

저자의 다른 논문

참고문헌 (21)

  1. Etchegaray, J. P. and M. Inouye. 1999. Translational enhancement by an element downstream of the initiation codon in Escherichia coli. J. Biol. Chem. 274: 10079- 10085 
  2. Kim, M. D., K.-N. Kim, and Y. J. Choi. 2003. Enhanced expression of $\beta$-xylosidase of Bacillus stearothermophilus No. 236 by change of translational inhibition codon in Escherichia coli and Bacillus subtilis. J. Microbiol. Biotechnol. 13: 584-590 
  3. Lim, D. and W. K. Maas. 1989. Reverse transcriptasedependent synthesis of a covalently linked, branched DNARNA compound in E. coli B. Cell 56: 891-904 
  4. Sambrook, J., E. F. Fritsch, and T. Maniatis. 1989. Molecular Cloning: A Laboratory Manual. 2nd Ed. Cold Spring Harbor Laboratory Press. Cold Spring Harbor, NY. U.S.A 
  5. Lim, D. and W. K. Maas. 1989. Reverse transcriptase in bacteria. Mol. Microbiol. 3: 1141-1144 
  6. Oh, K.-S., Y.-S. Park, and H.-C. Sung. 2002. Expression and purification of an ACE-inhibitory peptide multimer from synthetic DNA in Escherichia coli. J. Microbiol. Biotechnol. 12: 59-64 
  7. Sorensen, M. A., C. G. Kurland, and S. Pedersen. 1989. Codon usage determines translation rate in Escherichia coli. J. Mol. Biol. 207: 365-377 
  8. Lee, J.-H., Y.-D. Cho, J. J. Choi, Y.-J. Lee, H.-S. Hoe, H.-K. Kim, and S.-T. Kwon. 2003. High-level expression in Escherichia coli of alkaline phosphatase from Thermus caldophilus GK24 and purification of the recombinant enzyme. J. Microbiol. Biotechnol. 13: 660-665 
  9. Lim, D. 1991. Structure of two retrons of Escherichia coli and their common chromosomal insertion site. Mol. Microbiol. 5: 1863-1872 
  10. Kuroda, H. and P. Maliga. 2001. Sequences downstream of the translation initiation codon are important determinants of translation efficiency in chloroplasts. Plant Physiol. 125: 430-436 
  11. Lim, D., T. A. Gomes, and W. K. Maas. 1990. Distribution of msDNAs among serotypes of enteropathogenic Escherichia coli strains. Mol. Microbiol. 4: 1711-1714 
  12. Kim, K., D. W. Jeong, and D. Lim. 1997. A mutational study of the site-specific cleavage of EC83, a multicopy singlestranded DNA (msDNA): Nucleotides at the msDNA stem are important for its cleavage. J. Bacteriol. 179: 6518- 6521 
  13. Jeong, D. W., K. Kim, and D. Lim. 1997. Evidence for the complex formation between reverse transcriptase and multicopy single-stranded DNA in retron EC83. Mol. Cells 7: 347-351 
  14. Lim, D. 1992. Structure and biosynthesis of unbranched multicopy single-stranded DNA by reverse transcriptase in a clinical Escherichia coli isolate. Mol. Microbiol. 6: 3531- 3542 
  15. Temin, H. M. 1989. Retrons in bacteria. Nature 339: 254- 255 
  16. Furuichi, T., S. Inouye, and M. Inouye. 1987. Biosynthesis and structure of stable branched RNA covalently linked to the 5' end of multicopy single-stranded DNA of Stigmatella aurantiaca. Cell 48: 55-62 
  17. Maas, W. K., C. Wang, T. Lima, G. Zubay, and D. Lim. 1994. Multicopy single-stranded DNAs with mismatched base pairs are mutagenic in Escherichia coli. Mol. Microbiol. 14: 437-441 
  18. Sprengart, M. L., E. Fuchs, and A. G. Porter. 1996. The downstream box: An efficient and independent translation initiation signal in Escherichia coli. EMBO J. 15: 665-674 
  19. Etchegaray, J. P. and M. Inouye. 1999. A sequence downstream of the initiation codon is essential for cold shock induction of cspB of Escherichia coli. J. Bacteriol. 181: 5852-5854 
  20. Oh, Y. P., S. T. Jeong, D.-W. Kim, E.-C. Kim, and K.-H. Yoon. 2002. Simple purification of shiga toxin B chain from recombinant Escherichia coli. J. Microbiol. Biotechnol. 12: 986-988 
  21. Kim, J. S., H. Y. Seong, M. W. Kim, J. S. Ku, and S. Y. Choi. 2003. Effects of minor arginyl tRNA and isoleucyl tRNA on the expression of Clostridium botulinum neurotoxin light chain in Escherichia coli. J. Microbiol. Biotechnol. 13: 287- 291 

이 논문을 인용한 문헌 (0)

  1. 이 논문을 인용한 문헌 없음


원문 PDF 다운로드

  • ScienceON :
  • KCI :

원문 URL 링크

원문 PDF 파일 및 링크정보가 존재하지 않을 경우 KISTI DDS 시스템에서 제공하는 원문복사서비스를 사용할 수 있습니다. (원문복사서비스 안내 바로 가기)

상세조회 0건 원문조회 0건

DOI 인용 스타일