Gene Expression and Gut Microbiome Analysis in An. stephensi

For gene expression analysis in An. stephensi, total RNA was extracted from mosquitoes 24 hpi utilizing the TRIzol method (Sigma-Aldrich, China). Reverse transcription and quantitative PCR were performed as previously described [43 (link)]. The expression levels of target genes were normalized by the An. stephensi ribosomal gene S7. For detection of the abundance of gut microbiota, three midguts were pooled for DNA extraction. The levels of 16S rRNA gene were determined by quantitative PCR. The primers used for this study are listed in S2 Table.

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Feng Y., Chen L., Gao L., Dong L., Wen H., Song X., Luo F., Cheng G, & Wang J. (2021). Rapamycin inhibits pathogen transmission in mosquitoes by promoting immune activation. PLoS Pathogens, 17(2), e1009353.

Publication 2021

TRIzol method

Expression genes Gene Gene expression analysis Gut microbiota Mosquitoes Primers Reverse transcription Ribosomal Rrna gene Trizol

Corresponding Organization : Tsinghua University

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Variable analysis

independent variables

Time point (24 hpi)

dependent variables

Gene expression levels of target genes
Abundance of gut microbiota (16S rRNA gene levels)

control variables

An. stephensi ribosomal gene S7 (for gene expression normalization)

controls

Positive control: Not explicitly mentioned.
Negative control: Not explicitly mentioned.

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