Korean Journal of Microbiology (미생물학회지)

The Microbiological Society of Korea (한국미생물학회)
권호 표지

Autonomously Mitochondrial Replicating Sequence of Aspergillus nidulans

Aspergillus nidulans mtDNA의 자가복제절편

  • 장승환 (전북대 유전공학연구소 기초과학연구소 자연과학대학 생물과학부) ;
  • 한동민 (원광대 분자생물학과) ;
  • 장광엽 (전북대 유전공학연구소 기초과학연구소 자연과학대학 생물과학부)
  • Published : 1999.09.01
Download PDF
⟨ Previous Next ⟩

Abstract

We isolated the ANRI fragment from Aspergillus nidulons that could autononlously replicate and enhance transformation efficiency about $10^4$ fold compared lo the integrative vector in Saccha,omgcer cerevisioe. In A. nidulans recombinant plasmid pLJ16-4.5 which carries the 4.5 kb EcoRI fragment of ANRI showed a 170-[old increase of transformation efficiency compared to the integrative vector pLJ16 and could be recovered from iransfonnants as an intact form. Estimated copy number of transforming plasmid pLJ16-4.5 was scored as 2 to 3 copies in transformed A. nidulans. Recoinbinant plasinid pILJ16-4.5 is inilotically unstable; being lost Irom 65% of aswual progeny of transformants on selective medium and 90% on complete medium. Southern analysis of transformant DNA showed that the pILJ16-4.5 is maintained in free form. The sequencing data showed that ANRl fragment was originated from mitochondiral DNA of A. nid~ilans and contained high AT content as much as 74.7%. One ARS consensus sequence (A/T)TTr4T(A/G)TTT(AiT). I I ARS-like sequence (agreement 10 of 11) and ABFl binding core consensus sequence (TCN7ACG). Also six gyrase binding core consensus sequence (YRTGNYNNY: y=C or T, R=A or G, N=A, G, C or T) of $\Phi$X174 and SV40 DNA and one b site (CACTTTACC) combining with gyrase in ColEl are shown. ANRl can be developed as a repl&ng plasinid for lransfoimation system in A. nirlulmis.

Aspergillus nidulans 의 DNA 로부터 Saccharomyces cerevisiae에서 스스로 복제가능하고, 형질전환율도 삽입벡터에 비해 10\sup 4\ 배정도 높여주는 절편을 분리한 바 있다. A. nidulans에서 ANRI 의 일부를 포함한 pILJ16-4.5는 삽입벡터인 pILJ16보다 170배 정도 높은 형질전환 효율을 보였으며, S.cerevisiae와 마찬가지로 plasmid 상태로 회수가능했다. A.nidulans 페소내에서 2-3 copy 정도로 염색체와는 별개로 존재하는 것으로 나타났으며, 재형질전환능력도 있었다. ANRI은 미토콘드리아의 DNA에서 유래한 절편으로 밝고, ARS 공통절편과 유사한 절편도 11곳에 존재한다. $\Phi$X174와 SV40 DNA에서 gyrase 가 결합하는 부위의 공통편인 YRTGNYNNY도 6곳에 존재하며, ColE1에서 gyrase가 결합하는 b site와 일치하는 절편도 하나 포함하고 있으며, ABF1 결합 부위이 공통절편과 유사한 절푠$TCN_7ACG$ 도 하나 포함하고 있다. 이를 토대로 ANRI은 A.nidulans의 형질전환 후 회수가 가능한 replicating vector 개발에 이용할 수 있다.

Keywords

References

  1. Nucl.Acid. Res. v.18 no.19 Characterization of a novel plasmid-like element in Neurospora crassa derived mostly from the mitochondrial DNA Almasan,A.;N.C. Mishira
  2. Gene v.36 Development of a high-frequency transforming vector for Aspergillus nidulans Ballance, D.J.;G. Turner
  3. Curr. Genet. v.10 Behaviour of recombinant plasmid in Aspergillus nidulans: structure and stability Barnes, D.E.;D.W. MacDonald
  4. Nature v.275 no.5676 Transformation of yeast by a replicating hybrid plasmid Beggs,J.D.
  5. Curr. Genet. v.13 Behaviour of a replicating mitochondrial DNA sequence from Aspergillus amstelodami in Saccharomyces cerevisiae and Aspergillus nidulans Beri, R. K.;E.L. Lewis;G. Turner
  6. Mol. Cell. Biol. v.10 no.2 ARS binding factor Ⅰof the yeast Saccharomyces cerevisiae binds to sequences in telomeric and nontelomeric autonomously replicating sequences Biswas,S. B.;E.E. Biswas
  7. Cell v.51 no.5 A yeast silencer contains sequences that can promote autonomous plasmid replication and transcription activation Brand, A.H.;G. Micklem;K. Nasmyth
  8. The Molecular and Cellular Biology of the yeast Saccharomyces; Genome Dynamics, Protein Systhesis. and Energetics. v.1 Broach, J.R.;J.R. Pringle;E.W. Jones
  9. Mol. Cell. Biol. v.8 no.1 Two DNA-binding factors recognize specific sequences at silencers, upstream activating sequences, autonomously replicating sequences, and telomerers in Saccharomyces cerevisiae Buchman, A.R.;W.J. Kimmerly;J. Rine;R.D. Kornberg
  10. Science v.236 no.4803 Cloning of large segements of exgenous DNA into yeast by means of artificial chromosome vectors Burke, D.T.;G.F. Carle;M.V. Olson
  11. Mol. Gen. Genet. v.196 The transformation of mycelial spheroplasts of Neurospora crassa and the attempted isolation of an autonomous replicator Buxton, F.P.;A. Radford
  12. Proc. Natl. Acad. Sci. USA v.76 Efficient transformation of Neurospora. crassa. by utilizing hybrid plasmid DNA Case,M.E.;M. Schweizer;S.R. Kushner;N.H. Giles
  13. Nature v.287 Isolation of a yeast centromere and construction of functional small circular chromsome Clarke, L.;J.Carbon
  14. Cell v.28 Replication of Animal Mitochondrial DNA Clayton,D.A.
  15. GENE v.32 The ori sequences of the mitochondrial genome of a wild-type yeast strain: number, location, orientation and structure de Zamaroczy, M.;G. Faugeron-Fonty;G. Baldacci;R. Gourost;G. Bernardi
  16. Proc. Natl. Acad. Sci. USA v.85 Purification of a yeast protein that binds to origins to DNA replication and transcriptional silencer Diffley, J.F.X.;B. Stillman
  17. Nucl. Acid. Res. v.16 no.15 Identification of two factors which bind to the upstream sequences of a number of genes coding for mitochondrial proteins and to genetic elements important for cell division in yeast Dorsman, J.C.;W.C. van Heeswijl;L.A. Grivell
  18. Proc. Natl. Acad. Sci. USA. v.85 Specific interaction between a Saccharomyces cerevisiae protein and a DNA element associated with certain autonomously replicating sequences. Eisenberg, S.;C. Civalier;B.-K. Tye
  19. GENE v.98 An autonomously replicating plasmid transforms Aspergillus nidulans at high frequency Gems, D.;J.L. Johnstone;A.J. Clutterbuck
  20. Genetics v.121 Recombination and Replication of Plasmid-Like Derivatives of a Short Section of the Mitochondrial Chromosome of Neurospora crassa Cross, S.R.;P.H. Levine;S.Metzger;G. Glaser
  21. EMBO J. v.8 no.10 Isolation and DNA-binding characteristics of a protein involved in transcription activation of two divergently transcribed, essential yeast genes. Halfter, H.U.;M. Iler;E.-L. Winnacker;D.Gallwitz
  22. Mol. Cell. Biol. v.8 no.10 Constitutive Transcription of Yeast Ribosomal Protein Gene TCM1 Is Promoted by Uncommon cis-and trans-Acting Elements Hamil, K.G.;G.N.Hong; Fried H.M.
  23. Proc. Natl. Acad. Sic. USA v.80 no.4 Chimeric plasmid that replicates autonomously in both Esherichia coli and Neurospora crassa Hughes, K.;M.E. Case;R. Geever;D. Vapnek;N.H. Giles
  24. J. Microbiol. v.37 no.2 Isolation of an Autonomously Replicating DNA Sequence from Aspergillus nidulans Jang, S.H.;K.Y. Jahng
  25. Molecular cloning: a laboratory manual(2nd edition) Sambrook, J.;E.F. Fritch;T. Maniatis
  26. Proc. Natl. Acad. Sic. USA v.78 no.3 Contacts between DNA gyrase and its binding site on DNA: Feautures of symmetry and asymmetry revealed by protection from nucleases Morrison, A.;N.R. Cozzarelli
  27. J. Cell Biol. v.104 Regulation of the mRNA levels of nimA, a gene for the G₂-M transition on Aspergillus nidulnas Osmani, S.A.;G.S. May;N.R.Morris
  28. Curr. Genet. v.9 no.5 Plasmid recovery from transformants and the isolation of chromosomal DNA segments improving plasmid replication in Neurospora crassa Paietta, J.;G.A. Marzluf
  29. Proc. Natl. Acad. Sci. USA v.74 DNA sequencing with chain-terminating inhibitors Sanger, F.;S. Nicklen;A.R. Coulson
  30. J. Biol. Chem. v.265 no.25 ABF1 Binding Sites in Yeast Polymerase Genes. Seta, F.D.;I. Treich;J. -M. Buhler;A. Sentenac
  31. Proc. Natl. Acad. Sci. USA v.79 no.11 Replication and expression of a bacterial-mitochondrial hybrid plasmid in the fungus Podospora anserina Stahl, U.;P. Tudzynski;K. Esser
  32. J. Mol. Biol. v.158 no.2 Centromeric DNA from Sacchromyces cerevisiae Stinchomb, D.T.;R.W. Davis
  33. Nature v.282 Isolation and characterization of a yeast chromosomal replicator Stinchomb, D.T.;K. Struhl;R.W. Davis
  34. Proc. Natl. Acad. Sic. USA v.80 no.4 Construction of shuttle vector for the filamentous fungus Neuropora crassa Stohl, L.;A.M. Lambowitz
  35. Proc. Natl. Acad. Sci. USA v.76 High-frequency transformation of yeast: Autonomous replication of hybrid DNA molecules Struhl, K.;D.T. Stinchomb;S. Scherer;R.W. Davis
  36. Cell v.29 no.1 Cloning yeast telomeres on linear plasmid vectors Szostak, J.W.;E.H. Blackburn
  37. J. Biol. Chem. v.263 no.33 Purification and Characterization Proteins That Bind to Yeast ARSs. Sweder, K.S.;P.R. Rhode;J.L. Campbell
  38. Proc. Natl. Acad. Sci. USA v.87 A DNA replication enhancer in Saccharomyces cerevisiae Walker, S. S.;S.C. Francesconi;S. Eisenberg
  39. Proc. Natl. Acad. Sci. USA. v.81 Transformation of Aspergillus nidulans by using trpC plasmid Yelton, M. M.;J.E. Hamer;W.E. Timberlake
  40. Schizosaccaromyces pombe. Chromosoma v.103 Comparison of the two major ARS elements of the ura4 replication origin region with other ARS elements in the fission yeast Zhu;J.D.L. Carlson;D.D. Dubey;K. Sharma;J.A. Huberman