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Biolistic Transformation of Brugia Malayi

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 940))

Abstract

Biolistics has become a versatile tool for direct gene transfer to various cell and tissue types. Following its successful use on the parasitic nematode Ascaris suum, we developed and evaluated biolistics in the transfection of the model filarial parasite Brugia malayi. Biolistics was proven to be an efficient strategy for transfection of all life stages of the parasite and paved the way for studies on elements essential for promoter function and gene regulation of filarial parasites. Here we present a biolistics protocol for the transfection of B. malayi based on the Biolistics PDS 1000/He system and gold microcarriers.

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References

  1. Wilm T et al (1999) Ballistic transformation of caenorhabditis elegans. Gene 229:31–35

    Article  PubMed  CAS  Google Scholar 

  2. Jackstadt P et al (1999) Transformation of nematodes via ballistic DNA transfer. Mol Biochem Parasitol 103:261–266

    Article  PubMed  CAS  Google Scholar 

  3. Davis RE et al (1999) Transient expression of DNA and RNA in parasitic helminths by using particle bombardment. Proc Natl Acad Sci USA 96:8687–8692

    Article  PubMed  CAS  Google Scholar 

  4. Higazi TB et al (2002) Brugia malayi: transient transfection by microinjection and particle bombardment. Exp Parasitol 100(2):95–102

    Article  PubMed  CAS  Google Scholar 

  5. Shu L et al (2003) Analysis of the Brugia malayi HSP70 promoter using a homologous transient transfection system. Mol Biochem Parasitol 128(1):67–75

    Article  PubMed  CAS  Google Scholar 

  6. Higazi TB, Shu L, Unnasch TR (2004) Development and transfection of short-term primary cell cultures from Brugia malayi. Mol Biochem Parasitol 137(2):345–348

    Article  PubMed  CAS  Google Scholar 

  7. Higazi TB et al (2005) Identification of elements essential for transcription in Brugia malayi promoters. J Mol Biol 353:1–13

    Article  PubMed  CAS  Google Scholar 

  8. Liu C et al (2007) Sequences necessary for trans-splicing in transiently transfected Brugia malayi. Mol Biochem Parasitol 156:62–73

    Article  PubMed  CAS  Google Scholar 

  9. de Oliveira A, Katholi CR, Unnasch TR (2008) Characterization of the promoter of the Brugia malayi 12kDa small subunit ribosomal protein (RPS12) gene. Int J Parasitol 38:1111–1119

    Article  PubMed  Google Scholar 

  10. Liu C et al (2009) The splice leader addition domain represents an essential conserved motif for heterologous gene expression in B. malayi. Mol Biochem Parasitol 166:15–21

    Article  PubMed  CAS  Google Scholar 

  11. Liu C et al (2010) Functional analysis of putative operons in Brugia malayi. Int J Parasitol 40:63–71

    Article  PubMed  CAS  Google Scholar 

  12. Liu C, Chauhan C, Unnasch TR (2010) The role of local secondary structure in the function of the trans-splicing motif of Brugia malayi. Mol Biochem Parasitol 169:115–119

    Article  PubMed  CAS  Google Scholar 

  13. Tzertzinis G et al (2011) Molecular evidence for a functional ecdysone signaling system in Brugia malayi. PLoS Negl Trop Dis 4:e625

    Article  Google Scholar 

  14. Bailey M et al (2011) The role of polymorphisms in the spliced leader addition domain in determining promoter activity in Brugia malayi. Mol Biochem Parasitol 176:37–41

    Article  PubMed  CAS  Google Scholar 

  15. Hannon GJ et al (1990) Trans splicing of nematode pre-messenger RNA in vitro. Cell 61:1247–1255

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This work was supported by grants from US National Institutes of Health (project #R01-AI48562), the Edna McConnell Clark Foundation, and the Provost’s office of the University of Alabama at Birmingham (to TRU).

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Correspondence to Tarig B. Higazi .

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Higazi, T.B., Unnasch, T.R. (2013). Biolistic Transformation of Brugia Malayi . In: Sudowe, S., Reske-Kunz, A. (eds) Biolistic DNA Delivery. Methods in Molecular Biology, vol 940. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-110-3_9

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  • DOI: https://doi.org/10.1007/978-1-62703-110-3_9

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-109-7

  • Online ISBN: 978-1-62703-110-3

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