title: Molecular cloning and preliminary function study of iron responsive element binding protein 1 gene from cypermethrin-resistant Culex pipiens pallens.
abstract: BACKGROUND
  Insecticide resistance jeopardizes the control of mosquito populations and mosquito-borne disease control, which creates a major public health concern. Two-dimensional electrophoresis identified one protein segment with high sequence homology to part of Aedes aegypti iron-responsive element binding protein (IRE-BP).
  
  
  METHODS
  RT-PCR and RACE (rapid amplification of cDNA end) were used to clone a cDNA encoding full length IRE-BP 1. Real-time quantitative RT-PCR was used to evaluate the transcriptional level changes in the Cr-IRE strain Aedes aegypti compared to the susceptible strain of Cx. pipiens pallens. The expression profile of the gene was established in the mosquito life cycle. Methyl tritiated thymidine (3H-TdR) was used to observe the cypermethrin resistance changes in C6/36 cells containing the stably transfected IRE-BP 1 gene of Cx. pipiens pallens.
  
  
  RESULTS
  The complete sequence of iron responsive element binding protein 1 (IRE-BP 1) has been cloned from the cypermethrin-resistant strain of Culex pipiens pallens (Cr-IRE strain). Quantitative RT-PCR analysis indicated that the IRE-BP 1 transcription level was 6.7 times higher in the Cr-IRE strain than in the susceptible strain of 4th instar larvae. The IRE-BP 1 expression was also found to be consistently higher throughout the life cycle of the Cr-IRE strain. A protein of predicted size 109.4 kDa has been detected by Western blotting in IRE-BP 1-transfected mosquito C6/36 cells. These IRE-BP 1-transfected cells also showed enhanced cypermethrin resistance compared to null-transfected or plasmid vector-transfected cells as determined by 3H-TdR incorporation.
  
  
  CONCLUSIONS
  IRE-BP 1 is expressed at higher levels in the Cr-IRE strain, and may confer some insecticide resistance in Cx. pipiens pallens.
authors.affiliation: Department of Pathogenic Biology, Jining Medical University, Jining, Shandong Province, 272067, PR China. 1392144@163.com
authors.forename: Wenbin
authors.initials: W
authors.lastname: Tan
authors.forename: Xiao
authors.initials: X
authors.lastname: Wang
authors.forename: Peng
authors.initials: P
authors.lastname: Cheng
authors.forename: Lijuan
authors.initials: L
authors.lastname: Liu
authors.forename: Haifang
authors.initials: H
authors.lastname: Wang
authors.forename: Maoqing
authors.initials: M
authors.lastname: Gong
authors.forename: Xin
authors.initials: X
authors.lastname: Quan
authors.forename: Honggang
authors.initials: H
authors.lastname: Gao
authors.forename: Changliang
authors.initials: C
authors.lastname: Zhu
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D003465:Culex / Q000145:classification / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D005260:Female; D019476:Insect Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D007305:Insecticide Resistance; D007306:Insecticides / Q000494:pharmacology; D035925:Iron-Regulatory Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D010802:Phylogeny; D011722:Pyrethrins / Q000494:pharmacology*; D016415:Sequence Alignment
chemical_list: D019476:Insect Proteins; D007306:Insecticides; D035925:Iron-Regulatory Proteins; D011722:Pyrethrins; C017160:cypermethrin
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