scholarly journals Functional Characterization of Ecdysis Triggering Hormone Receptors (AgETHR-A and AgETHR-B) in the African Malaria Mosquito, Anopheles gambiae

2021 ◽  
Vol 12 ◽  
Author(s):  
Vikas Jindal ◽  
Yoonseong Park ◽  
Donghun Kim
Keyword(s):  
Heterologous Expression ◽  
Anopheles Gambiae ◽  
Transcription Initiation ◽  
Hormone Receptors ◽  
Chinese Hamster Ovary Cells ◽  
Expression System ◽  
Functional Characterization ◽  
Manual Annotation ◽  
Malaria Mosquito ◽  
Ecdysis Triggering Hormone

Insect ecdysis behavior, shedding off the old cuticle, is under the control of specific neuropeptides with the top command by the ecdysis triggering hormone (ETH). We characterized the ETH receptor (ETHR) of the malaria mosquito, Anopheles gambiae, by manual annotation of the NCBI gene (AGAP002881) and functional analysis, using a heterologous expression system in a mammalian cell line. The two splicing variants of ETHRs, ecdysis triggering hormone receptors (AgETHR-A and AgETHR-B), a conserved feature among insects, included of four (552 aa) and five exons (635 aa), respectively. The main feature of manual annotation of the receptor was a correction of N-terminal and exon-intron boundaries of an annotated gene (AGAP002881). Interestingly, the functional expression of the receptor in Chinese hamster ovary cells required modification of the transcription initiation site for mammalian Kozak consensus. In the calcium mobilization assay using the heterologous expression of each receptor, AgETHR-B showed a higher sensitivity to AgETH-1 (28 times) and AgETH-2 (320 times) than AgETHR-A. The AgETHRs showed specificity only to the ETH group of peptides but not to other groups carrying the C-termini motifs as PRXamide, such as pyrokinin1/DH and pyrokinin2/PBAN. Ecdysis triggering hormone receptors (AgETHR-B) responded to different ETH variants of other insect species more promiscuously than AgETHR-A.

scholarly journals A viral over-expression system for the major malaria mosquito Anopheles gambiae

2014 ◽  
Vol 4 (1) ◽  
Author(s):  
Yasutsugu Suzuki ◽  
Guodong Niu ◽  
Grant L. Hughes ◽  
Jason L. Rasgon
Keyword(s):  
Anopheles Gambiae ◽  
Expression System ◽  
Malaria Mosquito ◽  
Over Expression ◽  
Mosquito Anopheles Gambiae

Antibodies against the cystic fibrosis transmembrane regulator

1992 ◽  
Vol 262 (2) ◽  
pp. C396-C404 ◽  
Author(s):  
C. M. Fuller ◽  
M. B. Howard ◽  
D. M. Bedwell ◽  
R. A. Frizzell ◽  
D. J. Benos
Keyword(s):  
Cystic Fibrosis ◽  
Serum Proteins ◽  
Polyclonal Antibodies ◽  
Chinese Hamster Ovary Cells ◽  
Expression System ◽  
Functional Characterization ◽  
Chinese Hamster ◽  
Cystic Fibrosis Transmembrane Regulator ◽  
Cftr Protein ◽  
Cystic Fibrosis Transmembrane

Rabbit polyclonal antibodies have been raised against high-performance liquid chromatography purified synthetic peptides corresponding to two discrete regions of the cystic fibrosis transmembrane regulator (CFTR) protein: the R-domain (residues 785-796) and the extreme COOH-terminus (residues 1467-1480). Antibodies (Ab) to each of these peptides were affinity purified either by passage over a peptide-derivatized agarose matrix (Ab 785) to produce monospecific polyclonal antibodies or by protein A affinity chromatography to purify the immunoglobulin G1 fraction free of other serum proteins (Ab 1467). These antibodies recognize a candidate CFTR protein in the colonic cell line T84, as determined by Western blot analysis and by immunoprecipitation and labeling of the precipitate with [gamma-32P]ATP in the presence of protein kinase A. Both antibodies precipitated CFTR-related polypeptides from four mammalian cell types (HeLa, Bsc-40, HEp-2, and Chinese hamster ovary cells) transfected with the full-length CFTR cDNA clone using a vaccinia T7 protein expression system. Similar results were observed using a yeast CFTR expression system. In each case the Mr values of the bands observed were consistent with that expected for the CFTR protein. These antibodies should be useful probes for the immunocytochemical localization, immunoaffinity purification, and ultimately the functional characterization of the CFTR protein.

scholarly journals Harnessing a Transient Gene Expression System in Nicotiana benthamiana to Explore Plant Agrochemical Transporters

Plants ◽  
2021 ◽  
Vol 10 (3) ◽  
pp. 524
Author(s):  
Bingqi Wu ◽  
Zhiting Chen ◽  
Xiaohui Xu ◽  
Ronghua Chen ◽  
Siwei Wang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Transient Expression ◽  
Nicotiana Benthamiana ◽  
Heterologous Gene Expression ◽  
Expression System ◽  
Functional Characterization ◽  
Transient Gene Expression ◽  
High Mobility ◽  
Gene Expression System

Functional characterization of plant agrichemical transporters provided an opportunity to discover molecules that have a high mobility in plants and have the potential to increase the amount of pesticides reaching damage sites. Agrobacterium-mediated transient expression in tobacco is simple and fast, and its protein expression efficiency is high; this system is generally used to mediate heterologous gene expression. In this article, transient expression of tobacco nicotine uptake permease (NtNUP1) and rice polyamine uptake transporter 1 (OsPUT1) in Nicotiana benthamiana was performed to investigate whether this system is useful as a platform for studying the interactions between plant transporters and pesticides. The results showed that NtNUP1 increases nicotine uptake in N. benthamiana foliar discs and protoplasts, indicating that this transient gene expression system is feasible for studying gene function. Moreover, yeast expression of OsPUT1 apparently increases methomyl uptake. Overall, this method of constructing a transient gene expression system is useful for improving the efficiency of analyzing the functions of plant heterologous transporter-encoding genes and revealed that this system can be further used to study the functions of transporters and pesticides, especially their interactions.

scholarly journals Low-resolution genome map of the malaria mosquito Anopheles gambiae.

1991 ◽  
Vol 88 (24) ◽  
pp. 11187-11191 ◽  
Author(s):  
L. B. Zheng ◽  
R. D. Saunders ◽  
D. Fortini ◽  
A. della Torre ◽  
M. Coluzzi ◽  
...  
Keyword(s):  
Anopheles Gambiae ◽  
Malaria Mosquito ◽  
Low Resolution ◽  
Genome Map ◽  
Mosquito Anopheles Gambiae

scholarly journals Odorant reception in the malaria mosquito Anopheles gambiae

Nature ◽  
2010 ◽  
Vol 464 (7285) ◽  
pp. 66-71 ◽  
Author(s):  
Allison F. Carey ◽  
Guirong Wang ◽  
Chih-Ying Su ◽  
Laurence J. Zwiebel ◽  
John R. Carlson
Keyword(s):  
Anopheles Gambiae ◽  
Malaria Mosquito ◽  
Mosquito Anopheles Gambiae
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