| Neural peptide receptor (neuropeptide receptor, npr) is one type of G protein-coupled receptors (GPCRs), which can receive a variety of neuropeptides and intracellular signal including the FLPs and transmit them to the cells. This study focus on two kinds of southern root-knot nematode (Meloidogyne incognita Chitwood) neural peptide receptor gene:Mi-npr-1 and Mi-npr-3, the major jobs are as follow:1. According to the prediction of bio informatics and connected with the use of RACE technique, came out the open reading frame (ORF) of npr-1 and npr-3 through PCR. The full sequence of npr-1 is 1254bp with the GenBank number FJ456886. While the npr-3 is 1035bp with the number of GQ497771. The bioinformatics analysis showed that the amino acid sequence of NPR-1 and NPR-3 protein contain 7 transmemerbrane structures and this is the typical characteristics of G protein-coupled receptors.2. Having tried to use both prokaryotic and eukaryote expression of npr-1 and npr-3 and successfully cloned the target gene into the expression vector of pET-30a and pPIK9K. But In the induced expression of prokaryotic and eukaryotic, there was no finding of target protein in the SDS-PAGE electrophoresis.3. After the Analysis of npr-1 and npr-3 expressed in eggs, J2 and adult nematodes, the research indicated that they were expressed in the three stages of nematode development, which indicated that the npr-1 and npr-3 gene might playing an important role in the life cycle of M. incognita.4. Soaking treatment using J2 of M. incognita to carry out interference effects in vitro validation. The result shows that there was a significant difference in the root-knots number. There did not have difference among CK1 and CK2. Real-time quantitative RT-PCR analysis showed that lower transcript abundance for the targeted mRNAs than control. All the above indicated that npr-1 plays an important role in the development and parasitism of nematodes. |
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