title: RNA interference to reveal roles of β-N-acetylglucosaminidase gene during molting process in Locusta migratoria.
abstract: β-N-acetylglucosaminidases are crucial enzymes involved in chitin degradation in insects. We identified a β-N-acetylglucosaminidase gene (LmNAG1) from Locusta migratoria. The full-length complementary DNA (cDNA) of LmNAG1 consists of 2 667 nucleotides, including an open reading frame (ORF) of 1 845 nucleotides encoding 614 amino acid residues, and 233- and 589-nucleotide non-coding regions at the 5'- and 3'-ends, respectively. Phylogenetic analysis grouped the cDNA-deduced LmNAG1 protein with the enzymatically characterized β-N-acetylglucosaminidases in group I. Analyses of stage- and tissue-dependent expression patterns of LmNAG1 were carried out by real-time quantitative polymerase chain reaction. Our results showed that LmNAG1 transcript level in the integument was significantly high in the last 2 days of the fourth and fifth instar nymphs. LmNAG1 was highly expressed in foregut and hindgut. RNA interference of LmNAG1 resulted in an effective silence of the gene and a significantly reduced total LmNAG enzyme activity at 48 and 72 h after the injection of LmNAG1 double-stranded RNA (dsRNA). As compared with the control nymphs injected with GFP dsRNA, 50% of the dsLmNAG1-injected nymphs were not able to molt successfully and eventually died. Our results suggest that LmNAG1 plays an essential role in molting process of L. migratoria.
authors: Research Institute of Applied Biology, Shanxi University, Taiyuan.
authors: Shuo
authors: S
authors: Rong
authors: Da-Qi
authors: DQ
authors: Li
authors: Xue-Yao
authors: XY
authors: Zhang
authors: Sheng
authors: S
authors: Li
authors: Kun Yan
authors: KY
authors: Zhu
authors: Ya-Ping
authors: YP
authors: Guo
authors: En-Bo
authors: EB
authors: Ma
authors: Jian-Zhen
authors: JZ
authors: Zhang
mesh_terms: D000118:Acetylglucosaminidase / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D019476:Insect Proteins / Q000235:genetics* / Q000378:metabolism; D048188:Locusta migratoria / Q000145:classification / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D008969:Molecular Sequence Data; D018978:Molting; D010802:Phylogeny; D034622:RNA Interference
chemical_list: D019476:Insect Proteins; D000118:Acetylglucosaminidase
keywords: Locusta migratoria; RNA interference; chitin degradation; β-N-acetylglucosaminidase