title: Molecular cloning and characterization of a ryanodine receptor gene in brown planthopper (BPH), Nilaparvata lugens (Stål).
abstract: BACKGROUND
  Ryanodine receptors (RyRs) are a distinct class of intracellular calcium (Ca(2+)) release channel. The recent discovery of diamide insecticides has prompted studies on insect RyRs. However, information about the structure and function of insect RyRs is still limited. In this study, we isolated and characterized a full-length RyR cDNA (named NlRyR) from the brown planthopper, Nilaparvata lugens (Stål) (Homoptera: Delphacidae), a serious rice pest throughout Asia.
  
  
  RESULTS
  The composite NlRyR gene contains an open reading frame of 15 423 bp encoding a protein of 5140 amino acid residues, which shares high sequence identity (78-81%) with other insect homologues, except for two regions (IDR1: 4379-4732; IDR2: 1307-1529) with markedly low identity (44-48 and 38-41%, respectively). All hallmarks of the RyR proteins are conserved in the NlRyR protein, including the RyR domain as well as mannosyltransferase, IP3 R and RyR (pfam02815) (MIR) and RyR and IP3 R homology (pfam01365) (RIH) domains. Expression analysis of NlRyR revealed significant differences in mRNA expression levels among N. lugens developmental stages. Furthermore, three alternative splicing sites were identified in NlRyR, one of which forms the mutually exclusive exons A/B and is conserved in various insect species. Diagnostic PCR assays showed that the splice variant containing exon A was predominantly detected in all developmental stages.
  
  
  CONCLUSIONS
  NlRyR may play an important role in the control of developmental processes of N. lugens. Alternative splicing may generate the functional diversity of NlRyR. The results provided the basis for further structural and functional characterization of NlRyR.
authors.affiliation: College of Horticulture and Plant Protection, Yangzhou University, Yangzhou, China.
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.forename: Zhijuan
authors.initials: Z
authors.lastname: Xie
authors.forename: Jingkun
authors.initials: J
authors.lastname: Gao
authors.forename: Yaping
authors.initials: Y
authors.lastname: Liu
authors.forename: Wenlong
authors.initials: W
authors.lastname: Wang
authors.forename: Li
authors.initials: L
authors.lastname: Huang
authors.forename: Jianjun
authors.initials: J
authors.lastname: Wang
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D005091:Exons; D005260:Female; D005786:Gene Expression Regulation; D006430:Hemiptera / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D008297:Male; D008969:Molecular Sequence Data; D009758:Nymph / Q000235:genetics / Q000378:metabolism; D016366:Open Reading Frames; D010063:Ovum / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D019837:Ryanodine Receptor Calcium Release Channel / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D016415:Sequence Alignment
chemical_list: D018076:DNA, Complementary; D019476:Insect Proteins; D012333:RNA, Messenger; D019837:Ryanodine Receptor Calcium Release Channel
keywords: Nilaparvata lugens; alternative splicing; brown planthopper; calcium release channel; qRT-PCR; ryanodine receptor