Construction of the Suppression Subtractive Library of Candida Shehatae
|School||Nanjing Forestry University|
|Keywords||Suppression subtractive library Candida shehatae yeast Xylose fermentation Ethanol|
This is the first use of suppression subtractive hybridization , the successful establishment of the pentose metabolism typical strains of Candida shehatae yeast (Candida shehatae) hexose and pentose metabolic suppression subtractive library , the main results are as follows : xylose and glucose carbon source culture the C. shehatae, the the TRIZOL method of extraction of total RNA and mRNA purification kit , purified mRNA . Using suppression subtractive library construction kit to build the the C. shehatae suppression subtractive library , including regular library and anti- library . To identify the quality of the library were selected , the part of the differences in the anti - two libraries fragment gene sequencing , which is library 1000 ESTs were determined anti- library 500 ESTs were measured . Using Phred / Phrap / Consed package spliced ??the measured differences EST respectively Genebank Nt Database, Genebank Nr Database, SWlSSPROT Database using the BLAST software for sequence similarity comparison and classification GO classification , in contrast to gene . Analysis of the partial differential gene library using real-time fluorescence quantitative analysis method , the results further show that the better quality of the two libraries obtained in this research . The success of this study has been part of the gene of Candida shehatae yeast in different carbon sources cultured the part differential gene expression information and the flow associated with xylose metabolism , pentose yeast sugar metabolism at the transcriptional level functional genes conducted a preliminary study , the results of this study for microbial molecular metabolism of xylose metabolism , the level of transcription regulation and recombinant strains and other related research provides a good technical platform , bioinformatics dig for xylose fermentation ethanol production system to provide theoretical guidance to lay the theoretical foundation for the research and development of cellulosic ethanol .