Cloning And Expression Of Schizosaccharomyces Pombe Cdc2Gene In Microalgae

In this thesis, we use Microbial molecular biology and genetic engineering methods to study Schizosaccharomyces pombe cdc2gene cloning and expression of energy microalgae. In order to obtain a high oil content, a fast growing engineering microalgae. Provides certain experimental and theoretical basis for the construction of engineered microalgae.In this thesis,we take the use of quartz sand broken cell extraction, lysate extraction method and yeast genomic DNA extraction reagent box method to extract the genome DNA of S. pombe, by contrast found using yeast genomic DNA extraction reagent box method to extract the S. pombe genome DNA quality best.This paper takes use of the technology of PCR, getting the1189bp gene of cdc2from the cloning process of schizosaccharomyces pombe genome.Contrast the schizosaccharomyc-es pombe cdc2gene with schizosaccharomyces pombe cdc2gene which was reported in NCBI website, and the performance of BLAST sequence alignment shows that they have97%homology.Connect the schizosaccharomyces pombe cdc2gene with expression vector pBI121,constructing recombinant expression of pBI121-cdc2,and then use Electricity conversion leading the recombinant vector into navicula sp. cells, finally shape a navicula sp.of engineering microalgae.By drawing growth curve of navicula sp. of engineering microalgae and transformation of wild type navicula sp. we can see that the growth speed of successful transgenic navicula sp.engineering micro algae prominently increases which is about2times of wild type navicula sp.