摘要
Fungi play a significant role in biology-related domains, and with the molecular biology technology advancing, identification of fungi at molecular level and verification of genetic transformant has become the necessary step of test. By far, however, there is no ideal high-throughput molecular identification method. In this paper, a high-throughput device was designed, and a novel PCR-mediated molecular identification method suitable for the device was developed. Through cloning of ccllulase encoding genes and regulatory genes on the genome of Trichoderma reesei, cloning of promoter of phesphoglycerate kinase gene 1 and xylanase encoding genes on the genome and expression vector of Saccharomyces cerevisiae, and ITS sequences cloning and RAPD analysis of T. reesei, S. cerevisiae, Penicil- lium oxalicum, Rhizopus stolonifer, Aspergillus niger, A. carbonarius and A. japonicas, the method turned out to be an effective one with wide application potential. The establishment of the method has worked out the bottleneck of high-throughput molecular identification.
Fungi play a significant role in biology-related domains, and with the molecular biology technology advancing, identification of fungi at molecular level and verification of genetic transformant has become the necessary step of test. By far, however, there is no ideal high-throughput molecular identification method. In this paper, a high-throughput device was designed, and a novel PCR-mediated molecular identification method suitable for the device was developed. Through cloning of ccllulase encoding genes and regulatory genes on the genome of Trichoderma reesei, cloning of promoter of phesphoglycerate kinase gene 1 and xylanase encoding genes on the genome and expression vector of Saccharomyces cerevisiae, and ITS sequences cloning and RAPD analysis of T. reesei, S. cerevisiae, Penicil- lium oxalicum, Rhizopus stolonifer, Aspergillus niger, A. carbonarius and A. japonicas, the method turned out to be an effective one with wide application potential. The establishment of the method has worked out the bottleneck of high-throughput molecular identification.
基金
Supported by Applied Fundamental Research Project of Guangxi Department of Science and Technology(14125008-2-13)
