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Robust and regulatory expression of defensin A gene driven by vitellogenin promoter in transgenic Anopheles stephensi

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  • Entomology
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Chinese Science Bulletin

Abstract

The use of genetically modified mosquitoes to reduce or replace field populations is a new strategy to control mosquito-borne diseases. The precondition of the implementation of this strategy is the ability to manipulate the genome of mosquitoes and to induce specific expression of the effector molecules driven by a suitable promoter. The objective of this study is to evaluate the expression of defensin A gene of Anopheles sinensis under the control of a vitellogenin promoter in transgenic Anopheles stephensi. The regulatory region of Anopheles gambiae vitellogenin was cloned and subcloned into transfer vector pSLFa consisting of an expression cassette with defensin A coding sequence. Then, the expression cassette was transferred into transformation vector pBac[3xP3-DsRedafm] using Asc I digestion. The recombinant plasmid DNA of pBac[3xP3DsRed-AgVgT2-DefA] and helper plasmid DNA of phsp-pBac were micro-injected into embryos of An. stephensi. The positive transgenic mosquitoes were screened by observing specific red fluorescence in the eyes of G1 larvae. Southern blot analysis showed that a single-copy transgene integrated into the genome of An. stephensi. RT-PCR analysis showed that the defensin A gene expressed specifically in fat bodies of female mosquitoes after a blood meal. Interestingly, the mRNA of defensin A is more stable compared with that of the endogenous vitellogenin gene. After multiple blood meals, the expression of defensin A appeared as a reducible and non-cycling type, a crucial feature for its anti-pathogen effect. From data above, we concluded that the regulatory function of the Vg promoter and the expression of defensin A gene were relatively conserved in different species of anopheles mosquitoes. These molecules could be used as candidates in the development of genetically modified mosquitoes.

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Authors and Affiliations

  1. Institute of Tropical Diseases, School of Public Health and Tropical Medicine, Southern Medical University, Guangzhou, 510515, China

    Chen XiaoGuang, Zhang YaJing, Zheng XueLi & Wang ChunMei

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  1. Chen XiaoGuang
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  2. Zhang YaJing
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  3. Zheng XueLi
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  4. Wang ChunMei
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Correspondence to Chen XiaoGuang.

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Supported by the National Natural Science Foundation of China (Grant No. 30271162) and the Chinese Scholarship Council (Grant No. 2003365035)

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Chen, X., Zhang, Y., Zheng, X. et al. Robust and regulatory expression of defensin A gene driven by vitellogenin promoter in transgenic Anopheles stephensi . Chin. Sci. Bull. 52, 1964–1969 (2007). https://doi.org/10.1007/s11434-007-0292-z

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  • DOI: https://doi.org/10.1007/s11434-007-0292-z

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