KSBB Journal

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
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Analysis of Tissue Plasminogen Activator Expression using Pollen Culture in vitro

기내 화분배양을 이용한 Tissue Plasminogen Activator 발현분석

  • 박인혜 (대구가톨릭대학교 생명자원학부) ;
  • 박희성 (대구가톨릭대학교 생명자원학부)
  • Published : 2002.12.01
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Abstract

In an effort to use plant biotechnology for the production of biopharmaceuticals, pollens collected from lily (Lilium longiflorum) were grown in vitro and transformed with a PCR-amplified 1.7 kb cDNA encoding human tissue plasminogen activator (tPA) using Agrobacterium via a vacuum infiltration process. Western blotting showed that transgenic lily pollen tubes selected on kanamycin for 16 hrs expressed a tPA protein with a size similar to the human standard, suggesting their possible use as a disposable host for rapid foreign protein production.

백합(Lilium longiflorum)으로부터 수집한 화분에 대하여 기내배양, Agrobacterium 및 vacuum infiltration과정을 이용한 형질전환 그리고 kanamycin배지에서의 선별배양을 통하여 PCR에 의하여 증폭된 1.7 kb의 human tissue plasminogen activator(tPA) cDNA의 발현을 분석하였다. 16시간 정도 배양한 신장화분관의 western blotting결과, human standard와 유사한 크기로의 발현을 확인하였다. 이로써 백합화분은 신속한 단백질발현의 분석을 위한 일회성 생산숙주로서의 가능성을 제시하고 있다.

Keywords

References

  1. Curr. Opin. Biotech. v.11 Foreign protein production in plant tissue cultures Doran,P.M. https://doi.org/10.1016/S0958-1669(00)00086-0
  2. Curr. Opin. Biotech. v.12 Transgenic plants as protein factories Giddings,G. https://doi.org/10.1016/S0958-1669(00)00244-5
  3. Nature Biotech. v.18 Transgenic Plants as factories for biopharmaceuticals Giddings,G.;G.Allison;D.Brooks;A.Carter https://doi.org/10.1038/81132
  4. Curr. Opin. Biotech. v.12 Edible genetically modified microorganisms and plants for improved health Mercenier,A.;U.Wiedermann;H.Breiteneder https://doi.org/10.1016/S0958-1669(00)00255-X
  5. Plant Cell v.5 Male gametophyte development McCormick,S. https://doi.org/10.1105/tpc.5.10.1265
  6. Annu. Rev. Plant Physiol. Plant Mol. Biol. v.48 Pollen germination and tube growth Taylor,L.P.;P.K.Helper https://doi.org/10.1146/annurev.arplant.48.1.461
  7. Can. J. For. Res. v.28 Compatability of diferent pollination techniques with microprojectile bombardment of Norway spruce and Scots pine pollen Aronen,T.S.;T.O.Nikkanen;H.M.Haggman https://doi.org/10.1139/cjfr-28-1-79
  8. CR Acad. Sci. Paris Life Sci. v.316 In planta Agrobaterium-mediated gene transfer by infiltration of adult Arabidopsis thaliana plants Bethtold,N.;J.Ellis;G.Pellertier
  9. Plant Cell Rep. v.19 Transient gene expression in pine pollen tubes following particle bombardment Fernando,D.D.;J.N.Owens;S.Misra https://doi.org/10.1007/s002990050003
  10. Can. J. For. Res. v.27 Gene transfer by particle bombardment to Norway spruce and Scots pine pollen Haggman,H.M.;T.S.Aronen;T.O.Nikkanen https://doi.org/10.1139/cjfr-27-6-928
  11. Plant Sci. v.72 Transformation experiments by pipetting Agrobacterium into the spikelete of wheat(Triticum aestivum L.) Hess,D.;K.Dressler;R.Nimmrichter https://doi.org/10.1016/0168-9452(90)90087-5
  12. Plant J. v.2 Transformation of cereals via Agrobacterium and the pollen pathway; a critical assessment Langridge P.;R.Brettschneider;P.lazzeri;H.Lorz https://doi.org/10.1111/j.1365-313X.1992.00631.x
  13. Plant Mol. Biol. Rep. v.7 A simple method for the transformation of rice via the pollen-tuve pathway Luo,Z.X.;R.Wu https://doi.org/10.1007/BF02669248
  14. Plant Cell Rep. v.19 Vacuum infiltration of Petunia hybrida pollen with Agrobacterium tumefaciens to achieve plant transformation Tjokrokusumo,D.;T.Heinrich;S.Wylie;R.Potter;J.McComb https://doi.org/10.1007/s002990050009
  15. Sex. Plant Reprod. v.13 Pollen vegetative cell-specific expression of Bra rl: useful tool for observation of the vegetative nucleus and ientification of transgenic pollen by nuclear-targeted GFP Okada,T.;K.Toriyama https://doi.org/10.1007/s004970100074
  16. Sex. Plant Reprod. v.12 Comparison of transient and stable expression by a pollen-specific promoter: the transformation results do not always agree Hamilton,D.A.;Y.H.Schwarz;J.Rueda;J.P.Mascarenhas https://doi.org/10.1007/s004970050197
  17. Appl. Micro. Biotech. v.41 Extra- and intracellular amino acid concentrations in continuous Chinese hamster ovary cell culture Hansen H.A.;C.Emborg https://doi.org/10.1007/BF00178489
  18. Gene v.66 Amplified expression constructs for human tissue-type plasminogen activator in Chinese hamster ovary cells: instablilty in the absence of selective pressure Weidle,U.H.;P.Buckel;J.Wienberg
  19. Appl. Environ. Micro. v.64 Expression of active human tissue-type plasminogen activator in Escherichia coli Qiu,J.;J.R.Swartz;G.Georgiou
  20. Biotechnology v.9 Transgenic production of a variantof human tissue-type plasminogen activator in goat milk: generation of transgenic goats and analysis of expression Ebert,K.M.;J.P.Selgrath;P.Dittullio;J.Denman;T.E.Smith;M.A.Memom;J.E.Schindler;G.M.Monastersky;J.A.Vitale;K.Gordon https://doi.org/10.1038/nbt0991-835
  21. Mol. Biol. Med. v.3 Cloning of cDNA coding for tissue-type plasminogen activator and its expression in E. coli Harris,T.J.;T.Patel;F.A.Marston;S.Little;J.S.Emtage;G.Opdenakker;G.Volckaert;W.Rombauts;A.Billiau;P.DeSomer
  22. Kor. J. Genet. v.19 Inhibition of PKR phosphorylation in vtro by Lac-Z fused double-stranded RNA binding protein(RBF) produced from E. coli Park,H.S.;J.W.Choi
  23. Plant Physiol. v.116 Actin-bundling protein isolated from pollen tubes of lily Yokota,E.;K.Takahara;Y.Shimmen https://doi.org/10.1104/pp.116.4.1421
  24. Plant Molecular Biology Manual Binary vector An,G.;P.R.Ebert;A.Mitra;S.B.Ha;S.B.Gelvin(ed.);R.A.Schilperoort(ed.)