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The integrative expression of GUS gene driven by FCP promoter in the seaweed Laminaria japonica (Phaeophyta)

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Abstract

In this study, the background activity of β-glucuronidase (GUS) was analyzed histochemically and fluorometrically in the negative control of Laminaria japonica (Phaeophyta) thalli, showing low level of activity. GUS gene transformation without selectable gene in L. japonica was performed using four different promoters, i.e., Cauliflower mosaic virus 35S promoter (CaMV35S) from cauliflower mosaic virus, ubiquitin promoter (UBI) from maize, adenine-methyl transfer enzyme gene promoter (AMT) from virus in green alga Chlorella, and fucoxanthin chlorophyll a/c-binding protein gene promoter (FCP) from diatom Phaeodactylum tricornutum. The GUS transient activity was determined fluorometrically after bombarding sliced parthenogenetic sporophytes explants, and it was found that the activity resulting from CaMV35S and FCP promoters (in 114.3 and 80.6 pmol MU min−1 (mg protein)−1, respectively) was higher than for the other two promoters. The female gametophytes were bombarded and regenerated parthenogenetic sporophytes. FCP was the only promoter that resulted in detectable GUS chimeric expression activity during histochemical staining and polymerase chain reaction. Results of Southern blot showed that GUS gene was integrated with the L. japonica genome.

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References

  • Apt KE, Bhaya D, Grossman AR (1994) Characterization of genes encoding the light-harvesting proteins in diatoms: biogenesis of the fucoxanthin chlorophyll a/c protein complex. J Appl Phycol 6:225–230 doi:10.1007/BF02186075

    Article  CAS  Google Scholar 

  • Apt KE, Clendennen SK, Powers DA, Grossman AR (1995) The gene family encoding the fucoxanthin chlorophyll proteins from the brown alga Macrocystis pyrifera. Mol Gen Genet 246:455–464 doi:10.1007/BF00290449

    Article  PubMed  CAS  Google Scholar 

  • Bhaya D, Grossman AR (1993) Characterization of gene clusters encoding the fucoxanthin chlorophyll proteins of the diatom Phaeodactylum tricornutum. Nucleic Acids Res 21:4458–4466 doi:10.1093/nar/21.19.4458

    Article  PubMed  CAS  Google Scholar 

  • Bradford M (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254

    Article  Google Scholar 

  • Butler DM, Østgaard K, Boyen C, Evans LV, Jensen A, Kloareg B (1989) Isolation conditions for high yields of protoplasts from Laminaria saccharina and L. digitata (Phaeophyceae). J Exp Bot 40:1237–1246 doi:10.1093/jxb/40.11.1237

    Article  CAS  Google Scholar 

  • Caron L, Douady D, Quinet-Szely M, Goër S, Berkaloff C (1996) Gene structure of a chlorophyll a/c-binding protein from a brown alga: presence of an intron and phylogenetic implications. J Mol Evol 43:270–280 doi:10.1007/BF02338835

    Article  PubMed  CAS  Google Scholar 

  • Dai JX, Fang ZX (1976) Some preliminary observations on the parthenogenesis in Laminaria japonica Aresch. Acta Genet Sin 3:32–38 (in Chinese with English abstract)

    Google Scholar 

  • Douady D, Rousseau B, Caron L (1994) Fucoxanthin-chlorophyll a/c light-harvesting complexes of Laminaria saccharina: partial amino acid sequences and arrangement in thylakoid membranes. Biochemistry 33:3165–3170 doi:10.1021/bi00177a004

    Article  PubMed  CAS  Google Scholar 

  • Falciatore A, Casotti R, Leblanc C, Abrescia C, Bowler C (1999) Transformation of nonselectable reporter genes in marine diatoms. Mar Biotechnol 1:239–251 doi:10.1007/PL00011773

    Article  PubMed  CAS  Google Scholar 

  • Fang ZX (1983) A summary of the genetic studies of Laminaria japonica in China. In: Tseng CK (ed) Proceedings of the Joint China–U.S. Phycology Symposium. Science Press, Beijing, pp 123–136

    Google Scholar 

  • Goddijn OJM, Pen J (1995) Plants as bioreactors. Trends Biotechnol 13:379–387 doi:10.1016/S0167-7799(00)88985-4

    Article  CAS  Google Scholar 

  • Haq TA, Mason HS, Clements JD, Arntzen CJ (1995) Oral immunization with a recombinant bacterial antigen produced in transgenic plants. Science 268(5211):714–716 doi:10.1126/science.7732379

    Article  PubMed  CAS  Google Scholar 

  • Huang X, Weber JC, Hinson TK, Mathieson AC, Minocha SC (1996) Transient expression of the GUS reporter gene in the protoplasts and partially digested cells of Ulva lactuca L. (Chlorophyta). Bot Mar 39:467–474

    Article  CAS  Google Scholar 

  • Jefferson RA (1987) Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5:387–405 doi:10.1007/BF02667740

    Article  CAS  Google Scholar 

  • Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6(13):3901–3907

    PubMed  CAS  Google Scholar 

  • Jiang P, Qin S, Tseng CK (2002) Expression of hepatitis B surface antigen gene (HBsAg) in Laminaria japonica (Laminariales, Phaeophyta). Chin Sci Bull 47:1438–1440 doi:10.1360/02tb9317

    Article  CAS  Google Scholar 

  • Jiang P, Qin S, Tseng CK (2003) Expression of the lacZ reporter gene in sporophytes of the seaweed Laminaria japonica (Phaeophyceae) by gametophyte-targeted transformation. Plant Cell Rep 21:1211–1216 doi:10.1007/s00299-003-0645-2

    Article  PubMed  CAS  Google Scholar 

  • Kuang M, Wang SJ, Li Y, Shen DL, Tseng CK (1998) Transient expression of exogenous GUS gene in Porphyra yezoensis (Rhodophyta). Chin J Oceanol Limnol 16(Suppl):56–61 doi:10.1007/BF02849081

    Google Scholar 

  • Kübler JE, Minocha SC, Mathieson AC (1994) Transient expression of the GUS reporter gene in protoplast of Porphyra miniata (Rhodophyta). J Mar Biotechnol 1:165–169

    Google Scholar 

  • Kurtzman AM, Cheney DP (1991) Direct gene transfer and transient expression in a marine red alga using the biolistic method. J Phycol 27(Suppl):42

    Google Scholar 

  • Martin T, Whner RV, Hummel S, Willmitzer L, Fommer WB (1992) The GUS Gene Reporter system as a tool to study plant gene expression. In: Gallagher SR (ed) GUS Protocols: Using the GUS Gene as a Reporter of Gene Expression. Academic Press, San Diego, pp 23–43

    Google Scholar 

  • Matsumura W, Yasui H, Yamamoto H (2000) Mariculture of Laminaria japonica (Laminariales, Phaeophyceae) using protoplast regeneration. Phycol Res 48:169–176 doi:10.1111/j.1440-1835.2000.tb00213.x

    Article  Google Scholar 

  • Moffat AS (1995) Exploring transgenic plants as new vaccine source. Science 268(5211):658–660 doi:10.1126/science.7732373

    Article  PubMed  CAS  Google Scholar 

  • Qin S, Zhang J, Li WB, Wang XH, Tong S, Sun YR, Tseng CK (1994) Transient expression of GUS gene in Phaeophytes using Biolistic Particle Delivery System. Oceanol Limnol Sin 25:353–356 (in Chinese with English abstract)

    Google Scholar 

  • Qin S, Jiang P, Li XP, Tseng CK (1998a) The expression of lacZ in regenerated sporophytes of parthenogenetic Laminaria japonica. In: Menasveta P, Tanticharoen M (eds) Proceedings of the 2nd Asia-Pacific Marine Biotechnology Conference and 3rd Asia–Pacific Conference Algal Biotechnology. BIOTECH Press, Bangkok, pp 205–208

    Google Scholar 

  • Qin S, Jiang P, Li XP, Wang XH, Tseng CK (1998b) A transformation model for Laminaria japonica. Chin J Oceanol Limnol 16(Suppl):50–55 doi:10.1007/BF02849080

    Google Scholar 

  • Qin S, Sun GQ, Jiang P, Zou LH, Wu Y, Tseng CK (1999) Review of genetic engineering of Laminaria japonica (Laminariales, Phaeophyta) in China. Hydrobiologia 398/399:469–472 doi:10.1023/A:1017091629539

    Article  Google Scholar 

  • Qin S, Jiang P, Tseng CK (2005) Transforming kelp into a marine bioreactor. Trends Biotechnol 23(5):264–268 doi:10.1016/j.tibtech.2005.03.010

    Article  PubMed  CAS  Google Scholar 

  • Saga N, Sakai Y (1984) Isolation of protoplasts from Laminaria and Porphyra. Bull Jap Soc Sci Fish 50:1085

    Google Scholar 

  • Saunders GW (1993) Gel purification of red algal genomic DNA: an inexpensive and rapid method for the isolation of polymerase chain reaction-friendly DNA. J Phycol 29:251–254 doi:10.1111/j.0022-3646.1993.00251.x

    Article  CAS  Google Scholar 

  • Taylor MG, Vasil V, Vasil IK (1993) Enhanced GUS gene expression in cereal/grass cell suspensions and immature embryos using the maize ubiquitin-based plasmid pAHC25. Plant Cell Rep 12:491–495

    CAS  Google Scholar 

  • Tseng CK, Qin S (1991) Marine algal biotechnology in China: Present conditions and prospects. In: Proceeding of International Symposium on Biotechnology of Salt Ponds. Tianjin, pp. 61–68

  • Walden R, Wingender R (1995) Gene-transfer and plant-regeneration (techniques). Trends Biotechnol 13:324–331 doi:10.1016/S0167-7799(00)88976-3

    Article  CAS  Google Scholar 

  • Wang XH, Qin S, Li XP, Jiang P, Tseng CK (1998) High efficient induction of callus and regeneration of sporophytes of Laminaria japonica (Phaeophyta). Chin J Oceanol Limnol 16(Suppl):67–74 doi:10.1007/BF02849083

    Google Scholar 

Download references

Acknowledgements

This work was supported by the Key Innovative Project of Chinese Academy of Science (KZCX2-YW-209) and the CAS/SAFEA International Partnership Program for Creative Research Teams (Research and Applications of Marine Functional Genomics).

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Authors and Affiliations

  1. Institute of Oceanology, Chinese Academy of Sciences, 266071, Qingdao, People’s Republic of China

    Fuchao Li, Song Qin, Peng Jiang, Yun Wu & Wei Zhang

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  1. Fuchao Li
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  2. Song Qin
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  3. Peng Jiang
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  5. Wei Zhang
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Correspondence to Song Qin.

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Li, F., Qin, S., Jiang, P. et al. The integrative expression of GUS gene driven by FCP promoter in the seaweed Laminaria japonica (Phaeophyta). J Appl Phycol 21, 287–293 (2009). https://doi.org/10.1007/s10811-008-9366-9

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  • DOI: https://doi.org/10.1007/s10811-008-9366-9

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