| Comparing differentially expressed genes of different growth and development stages in the same species is theoretically and practically important, which can provide valuable information for growth, development and metabolic pathway study.Period from dicaryotic mycelium to primordium is a complicated development process for Pleurotus ostreatus. To date, few studies on the primordia formation mechanism at molecular level have been reported. In this study, cDNA libraries between dicaryotic mycelium and primordia stage of Pleurotus ostreatus, were constructed by suppression subtractive hybridization method, and the screening and analysis of differentially expressed genes of the two development stages will give help for the understanding of Pleurotus ostreatus development mechanism at the molecular level.In this study, forward and reverse SSH cDNA libraries were constructed using primordia of Pleurotus ostreatus Xin831as tester and dicaryotic mycelium as driver, or dicaryotic mycelium as tester and primordia as driver, respectively.2000clones were obtained from each library. In order to verify the quality of library,ten positive clones from forward and reverse library were sequenced,respectively.The main results are as follows:(1) The high quality double-stranded cDNAs of dicaryotic mycelium and primordia were synthesized using SMARTerTM PCR cDNA Synthesis Kit, which provided a foundation for the SSH cDNA library construction.(2) Forward and reverse SSH cDNA libraries were constructed. The inserted cDNA size between250-750bp of clones was confirmed through colony PCR.2000clones from forward and reverse libraries were obtained, respectively.(3) Ten clones were selected randomly and sequenced from each library to verify the quality of SSH cDNA library construction. Some genes involved in regulating primordia formation were found, which indicated that the forward and reverse subtractive libraies were well constrcted. |
PDF Full Text Request