Related Experiment Videos
Identification and characterization of Laodelphax striatellus (Insecta: Hemiptera: Delphacidae) neutral
Y Zhou1,2, X-W Lin1, M-A Begum1
1State Key Laboratory of Rice Biology; Ministry of Agriculture Key Lab of Molecular Biology of Crop Pathogens and Insects; and Institute of Insect Sciences, Zhejiang University, Hangzhou, Zhejiang, China.
Abstract:
The neutral sphingomyelinase (nSMase) 1 homologue gene LsSMase was cloned from Laodelphax striatellus, a direct sap-sucker and virus vector of gramineous plants, and expressed via a Bac to Bac baculovirus expression system. The LsSMase-enhanced green fluorescent protein fusion protein was located in the endoplasmic reticulum in a similar manner to mammalian nSMase 1. The biochemical properties of LsSMase were determined in detail. The optimal pH and temperature for recombinant LsSMase were 8 and 37 °C, respectively. LsSMase was an Mg2+ or Mn2+ dependent enzyme, but different concentration of each were needed. The activity of LsSMase was significantly stimulated by Ethylene glycol bis(2-aminoethyl ether)tetraacetic acid (EGTA), whereas it was inhibited by ethylenediaminetetraacetic acid. Millimolar concentrations of Zn2+ completely inhibited LsSMase. The reducing agents dithiothreitol and β-mercaptoethanol varied in their effects on activity. Phospholipids were not found to stimulate LsSMase.