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Marine Biotechnology

, Volume 8, Issue 5, pp 491–500 | Cite as

Discovery of the Genes in Response to White Spot Syndrome Virus (WSSV) Infection in Fenneropenaeus chinensis Through cDNA Microarray

  • Bing Wang
  • Fuhua Li
  • Bo Dong
  • Xiaojun Zhang
  • Chengsong Zhang
  • Jianhai XiangEmail author
Original Article

Abstract

We used microarray technology to study differentially expressed genes in white spot syndrome virus (WSSV)-infected shrimp. A total of 3136 cDNA targets, including 1578 unique genes from a cephalothorax cDNA library and 1536 cDNA clones from reverse and forward suppression subtractive hybridization (SSH) libraries of Fenneropenaeus chinensis, plus 14 negative and 8 blank control clones, were spotted onto a 18 × 18 mm area of NH2-modified glass slides. Gene expression patterns in the cephalothorax of shrimp at 6 h after WSSV injection and moribund shrimp naturally infected by WSSV were analyzed. A total of 105 elements on the arrays showed a similar regulation pattern in artificially infected shrimp and naturally infected moribund shrimp; parts of the results were confirmed by semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR). The up-regulated expression of immune-related genes, including heat shock proteins (HSP70 and HSP90), trehalose-phosphate synthase (TPS), ubiquitin C, and so forth, were observed when shrimp were challenged with WSSV. Genes including myosin LC2, ATP synthase A chain, and arginine kinase were found to be down-regulated after WSSV infection. The expression of housekeeping genes such as actin, elongation factor, and tubulin is not stable, and so these genes are not suitable as internal standards for semiquantitative RT-PCR when shrimp are challenged by WSSV. As a substitute, we found that triosephosphate isomerase (TPI) was an ideal candidate of interstandards in this situation.

Keywords

Expressed sequence tags Fenneropenaeus chinensis microarray subtractive suppression hybridization library white spot syndrome virus 

Notes

Acknowledgments

We thank Professor P. Natarajar, TWAS-UNESCO, and Dr. Ka Hou Chu, Chinese University of Hong Kong for their constructive comments on this manuscript. This work was supported by the Key Program of National Natural Science Foundation of China (30230280) and the National High Technology Research and Development Program (2005AA626014).

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Copyright information

© Springer Science+Business Media, Inc. 2006

Authors and Affiliations

  • Bing Wang
    • 1
    • 2
  • Fuhua Li
    • 1
  • Bo Dong
    • 1
  • Xiaojun Zhang
    • 1
  • Chengsong Zhang
    • 1
  • Jianhai Xiang
    • 1
    Email author
  1. 1.Institute of OceanologyChinese Academy of SciencesQingdaoPeople's Republic of China
  2. 2.Graduate SchoolChinese Academy of SciencesBeijingPeople's Republic of China

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