Cloning of a novel protease required for the molting of Locusta migratoria manilensis.
Molting is required for progression between larval stages in the life cycle of an insect. The essence of insect molting is the laying down of new cuticle followed by shedding of the old cuticle. Degradation and recycling of old cuticle are brought about by enzymes present in the molting fluid, which fills the space between the old and new cuticle. Here, we describe the cloning of a novel protease gene from Locusta migratoria manilensis, designated as Lm-TSP. The cDNA and its deduced protein sequences were deposited in GenBank (accession numbers EF081255 and ABN13876, respectively). Sequence analysis indicated that Lm-TSP belongs to the trypsin-like serine protease family. We show, by RNA interference (RNAi), that silencing of Lm-TSP leads to dramatic reductions in protease and cuticle-degrading activity of a molting fluid, which leads to molting defects from fourth-instar larvae (L4) to fifth-instar larvae (L5), and between L5 and adult stages. These observations suggest that Lm-TSP plays a critical role in L. migratoria manilensis ecdysis.
Wei
Zhang
Z
Genetic Engineering Research Center; School of Bioengineering, Chongqing University, Chongqing 400044, China.
Yin
Youping
Y
Zhang
Basheng
B
Wang
Zhongkang
Z
Peng
Guoxiong
G
Cao
Yueqing
Y
Xia
Yuxian
Y
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D018076:DNA, Complementary; D006110:Grasshoppers / Q000502:physiology*; D008969:Molecular Sequence Data; D018978:Molting / Q000502:physiology*; D010447:Peptide Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000502:physiology*; D034622:RNA Interference; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D010447:Peptide Hydrolases