use-recombinant-mosquito-densovirus-as-gene-delivery-vector-for

Use of a Recombinant Mosquito Densovirus As a Gene Delivery Vector for the Functional Analysis of Genes in Mosquito Larvae

In vivo microinjection is the most commonly used gene transfer technique for analyzing the gene functions in individual mosquitoes. However, this method ...

Southern Medical University

To identify antigens which may help evaluate the therapeutic effect of angiostrongyliasis from adult worm antigen of Angiostrongylus cantonensis.

[Screening and identification of therapeutic effect evaluation antigens of angiostrongyliasis].

To reconstitute a transactivation system in yeast (yeast model) for screening the pesticides acting on ecdysone metabolism route and eventually influencing the process of ecdysis.

[Construction of a yeast model for screening Aedes albopictus ecdysone agonist pesticides].

To construct the life cycle of Angiostrongylus cantonensis (A.cantonensis) in laboratory condition.

[Construction of the life cycle of Angiostrongylus cantonensis in laboratory].

This paper reports a severe case of eosinophilic meningoencephalitis after infection with Angiostrongylus cantonensis.

A severe eosinophilic meningoencephalitis caused by infection of Angiostrongylus cantonensis.

To investigate the effect of cinobufagin on nuclear factor-kappaB (NF-kappaB) pathway of liver cancer cell line HepG2.

[Effect of cinobufagin on nuclear factor-kappaB pathway in HepG2 cells].

To verify the miRNA in Aedes albopictus and characterize the expression profile of several miRNAs across all the life stages of Aedes albopictus.

[Isolation, identification and analysis of the expression profile of miRNAs in Aedes albopictus].

To express antibacterial peptide thanatin in the prokaryotic expression system and test its antibacterial activity.

[Antibacterial activity of recombinant thanatin expressed by E.coli ER2566].

To analyze and predict the structure and function of mosquito densovirus (MDV) nostructual protein1 (NS1).

[Bioinformatics analysis of mosquito densovirus nostructure protein NS1].

The Aedes aegypti densovirus (AeDNV) has previously shown potential in mosquito control. To improve its efficacy as a biopesticide, the gene for an excitatory insect-specific toxin from Buthus martensii Karsch (BmK IT1) was inserted into the AeDNV genome and cloned into pUCA plasmid. The coding sequence for green fluorescent protein was ligated to the C-terminus of the BmK IT1 gene as a screening marker. Recombinant and helper plasmids were cotransfected into C6/36 cells; wild-type viruses were the controls. The recombinant viruses were identified and quantified by real-time polymerase chain reaction and exposed to Ae. albopictus larvae for the evaluation of its bioinsecticidal activity. LT(50) and LD(50) bioassays showed that the recombinant AeDNV had stronger and faster pathogenic effects on Ae. albopictus than the wild-type virus. This is the first report on the recombinant AeDNA containing the insect-specific toxin, BmK IT1, which may be used to develop a novel type of insecticide.

A recombinant AeDNA containing the insect-specific toxin, BmK IT1, displayed an increasing pathogenicity on Aedes albopictus.

An artificial intron consisting of the 5'-donor site (from the first intron of the human beta-globin gene) and the 3'-acceptor site (from the intron of an immunoglobulin gene heavy chain variable region) was obtained with a splice overlap extension PCR and was then inserted in frame into the coding sequence of nostructural protein NS1 gene fused to GFP gene in a recombinant mosquito densovirus plasmid p7NS1-GFP. The constructed plasmid was named as p7NS1-Intron-GFP. The plasmid p7NS1-Intron-GFP was co transfected with the helper plasmid pUCA into C6/36 cells, then the packaged recombinant and wild type viruses were purified and recovered. The second-instars of Aedes albopictus larvae were exposed to recombinant and wild type virus mixed stock. The high level GFP expression in C6/36 cells and larvae was observed under fluorescence microscope, indicating that the inserted artificial intron exerted its normal function in self-splicing both in vitro and in vivo. This study laid a foundation for application of an artificial intron in insect cells and development of new strategy for genetic engineering technology of mosqtuito and its pathogens.

[Preliminary application of a mosquito densovirus-mediated artificial intron in vitro and in vive of mosquito].

To prepare a large amount of pure alive Schistosoma japonicum eggs, rabbit was infected with 2000 cercariae and its liver was taken aseptically 38-45 days after infection and homogenized. The homogenate was screened through different sieves(60, 120, 200, 300, 360 meshes per inch respectively), and washed with 1.2% NaCl. The eggs and leftover were then digested with 0.25% trypsin for 2 hours, sieved over 360 meshes per inch and washed with RPMI 1640 medium. The collected eggs reached to (95.1 +/- 6.4)% of live eggs, with a high efficiency.

Article	Year
[Screening and identification of therapeutic effect evaluation antigens of angiostrongyliasis]. Nan fang yi ke da xue xue bao = Journal of Southern Medical University\| PubMed ID: 16546727	2006
[Construction of a yeast model for screening Aedes albopictus ecdysone agonist pesticides]. Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases\| PubMed ID: 18441988	2007
[Construction of the life cycle of Angiostrongylus cantonensis in laboratory]. Nan fang yi ke da xue xue bao = Journal of Southern Medical University\| PubMed ID: 18495588	2008
A severe eosinophilic meningoencephalitis caused by infection of Angiostrongylus cantonensis. The American journal of tropical medicine and hygiene\| PubMed ID: 18840746	2008
[Effect of cinobufagin on nuclear factor-kappaB pathway in HepG2 cells]. Nan fang yi ke da xue xue bao = Journal of Southern Medical University\| PubMed ID: 20118004	2010
[Isolation, identification and analysis of the expression profile of miRNAs in Aedes albopictus]. Nan fang yi ke da xue xue bao = Journal of Southern Medical University\| PubMed ID: 20423824	2010
[Antibacterial activity of recombinant thanatin expressed by E.coli ER2566]. Nan fang yi ke da xue xue bao = Journal of Southern Medical University\| PubMed ID: 20501392	2010
[Bioinformatics analysis of mosquito densovirus nostructure protein NS1]. Zhonghua shi yan he lin chuang bing du xue za zhi = Zhonghua shiyan he linchuang bingduxue zazhi = Chinese journal of experimental and clinical virology\| PubMed ID: 20718357	2009
A recombinant AeDNA containing the insect-specific toxin, BmK IT1, displayed an increasing pathogenicity on Aedes albopictus. The American journal of tropical medicine and hygiene\| PubMed ID: 20810829	2010
[Preliminary application of a mosquito densovirus-mediated artificial intron in vitro and in vive of mosquito]. Bing du xue bao = Chinese journal of virology\| PubMed ID: 21043138	2010
[Improved preparation of pure alive eggs of Schistosoma japonicum]. Zhongguo ji sheng chong xue yu ji sheng chong bing za zhi = Chinese journal of parasitology & parasitic diseases\| PubMed ID: 24812826	2008

Jin-Bao Gu

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