Studies of the Parasite-Midgut Interaction Reveal Plasmodium Proteins Important for Malaria Transmission to Mosquitoes

Malaria transmission relies on parasite-mosquito midgut interaction. The interactive proteins are hypothesized to be ideal targets to block malaria transmission to mosquitoes. We chose 76 genes that contain signal peptide-coding regions and are upregulated and highly abundant at sexual stages. Forty-six of these candidate genes (60%) were cloned and expressed using the baculovirus expression system in insect cells. Six of them, e.g., PF3D7_0303900, PF3D7_0406200 (Pfs16), PF3D7_1204400 (Pfs37), PF3D7_1214800, PF3D7_1239400, and PF3D7_1472800 were discovered to interact with blood-fed mosquito midgut lysate. Previous works showed that among these interactive proteins, knockout the orthologs of Pfs37 or Pfs16 in P. berghei reduced oocysts in mosquitoes. Here we further found that anti-Pfs16 polyclonal antibody significantly inhibited P. falciparum transmission to Anopheles gambiae. Investigating these candidate proteins will improve our understanding of malaria transmission and discover new targets to break malaria transmission.

Download Full-text

Production, processing and partial purification of functional G protein βγ subunits in baculovirus-infected insect cells

Biochemical Journal ◽

10.1042/bj2860677 ◽

1992 ◽

Vol 286 (3) ◽

pp. 677-680 ◽

Cited By ~ 21

Author(s):

J D Robishaw ◽

V K Kalman ◽

K L Proulx

Keyword(s):

G Protein ◽

G Proteins ◽

Insect Cells ◽

Expression System ◽

Baculovirus Expression System ◽

Partial Purification ◽

Baculovirus Expression ◽

Gamma Subunits ◽

Infected Insect ◽

Beta 2

As a result of the inability to resolve the heterogeneous mixture of G protein beta gamma subunits present in tissues, it has not been possible to compare different beta gamma subunits of the G proteins in terms of their proposed roles in receptor-effector coupling. This study was undertaken to establish the utility of the baculovirus expression system in producing homogeneous beta gamma subunits of defined composition for the comparative analysis of these subunits in reconstitution systems. In this study we report the expression, and appropriate post-translational processing, of recombinant beta 2, gamma 2 and gamma 3 subunits. In addition, we show that the recombinant beta gamma subunits can be readily purified, and can functionally interact with the alpha subunits of the G proteins.

Download Full-text

Improvement of Baculovirus Expression System and Purification of IL-6 Protein Expressed in Insect Cells

Chinese Journal of Biotechnology ◽

10.1016/s1872-2075(06)60046-0 ◽

2006 ◽

Vol 22 (4) ◽

pp. 572-580 ◽

Cited By ~ 3

Author(s):

N YAO ◽

L YAO ◽

Y KAN ◽

W ZHOU ◽

Y QI

Keyword(s):

Insect Cells ◽

Expression System ◽

Baculovirus Expression System ◽

Baculovirus Expression

Download Full-text

A secretary bi-cistronic baculovirus expression system with improved production of the HA1 protein of H6 influenza virus in insect cells and Spodoptera litura larvae

Journal of Immunological Methods ◽

10.1016/j.jim.2018.06.001 ◽

2018 ◽

Vol 459 ◽

pp. 81-89 ◽

Cited By ~ 3

Author(s):

Ming-Shou Hsieh ◽

Jie-Long He ◽

Tzong-Yuan Wu ◽

Rong-Huay Juang

Keyword(s):

Influenza Virus ◽

Spodoptera Litura ◽

Insect Cells ◽

Expression System ◽

Baculovirus Expression System ◽

Baculovirus Expression

Download Full-text

Partial characterization of natural and recombinant human soluble CD23

Biochemical Journal ◽

10.1042/bj2860819 ◽

1992 ◽

Vol 286 (3) ◽

pp. 819-824 ◽

Cited By ~ 7

Author(s):

K Rose ◽

G Turcatti ◽

P Graber ◽

S Pochon ◽

P O Regamey ◽

...

Keyword(s):

Insect Cells ◽

Peptide Mapping ◽

Sequence Similarity ◽

Expression System ◽

Protein A ◽

Baculovirus Expression System ◽

Partial Characterization ◽

Disulphide Bonds ◽

Baculovirus Expression

The purification to homogeneity of an active soluble 25 kDa fragment of CD23, produced in insect cells using the baculovirus expression system, is described. Peptide mapping and analysis by Edman degradation and mass spectrometry permitted partial characterization of the protein. A total of 165 out of 172 residues, including N-terminal and C-terminal regions, were mapped. The positions of the two disulphide bonds in the IgE-binding region were also determined: residue 110 is joined to residue 124, and residue 42 to residue 133. Natural CD23 25 kDa fragment was also analysed and found to possess the same disulphide bond arrangement. These results extend the previously noted sequence similarity with lectins to elements of secondary structure.

Download Full-text