English (United Kingdom)

https://curated-unify.zendy.io/wp-json/zendy-region/v1/featured_content/oa?rat=en

https://curated-unify.zendy.io/wp-json/zendy-region/v1/highlighted_journal/

Global: Zendy Plus annual

Presents the access of premium content as premium feature

Premium Content

Presents the keyphrase highlighting as premium feature

Keyphrase Highlighting

Presents the summarisation as premium feature

Summarisation

Insights

Presents the pdf analysis as premium feature

PDF Analysis

Presents the zaia usage as premium feature

ZAIA

Global: Zendy Tools annual

Global: Zendy Free Plan

Global: Zendy Tools monthly

Global: Zendy Plus monthly

Trypsin modulating oostatic factor (TMOF), a decapeptide hormone synthesized by female mosquito ovaries, ganglia and the central nervous system of  Aedes aegypti , terminates trypsin biosynthesis in larvae, and blood-fed female mosquitoes. Earlier, TMOF was cloned and expressed as a single copy in  Chlorella dessicata  and in  Saccharomyces cerevisiae  cells as a potential larvicide. Here we report the use of a methylotrophic yeast cells,  Pichia pastoris , that efficiently express multi copies of heterologous proteins, that are readily ingested by mosquito larvae.  P. pastoris  was engineered using pPICZB (Invitrogen, CA, United States), and 2 genes:  gfp-tmf A and  tmf A inserted between  Kpn I and  Xba I in the multiple cloning site. The plasmid carries a strong AOXI promoter and  P. pastoris  KM71 and KM71H cells were transformed by homologous recombination. The synthesis of GFP-TMOF was followed using UV and clones were analyzed using southern and Northern blot analyses. Cloning  tmf A into KM71H and selection on high Zeocin concentration (2.0 mg/mL) identified a clone that carried 10 copies of  tmfA.  A comparison between a single and high copy (10 genes) insertions using Northern blot analyses showed that a  tmfA  transcript was highly expressed even after 120 h. SDS-PAGE analysis of KM71 cells transformed with  gfp-tmf A identified a protein band that ran at the expected  M   r   of 31 kDa. Enzyme Linked Immunoadsorbant Assay (ELISA) analysis of the recombinant cells showed that 1.65 × 10 8  and 8.27 × 10 7  cells produce 229 and 114 μM of TMOF, respectively, and caused 100% larval mortality when fed to groups of 5 larvae in 25 mL water. These results indicate that the recombinant  P. pastoris  cells could be used in the future in the marsh to control mosquito populations.

Biochemical and Molecular Characterization of Pichia pastoris Cells Expressing Multiple TMOF Genes (tmfA) for Mosquito Larval Control