同源重组敲除三角褐指藻基因的研究

The research of gene knockout via homologous recombination in Phaeodactylum tricornutum

摘要:

三角褐指藻是海洋硅藻研究的模式藻种，也是目前最受关注的微藻生物能源藻种之一，基因组测序已经完成，而其基因功能解析和基因工程调控有待进一步探索. 本文构建了三角褐指藻甘油激酶基因敲除载体，成功转化至三角褐指藻中，经筛选鉴定获得34个阳性转基因藻株；并进一步对三角褐指藻甘油激酶基因敲除转基因藻株的甘油激酶表达量和生长两方面进行分析，结果显示胞外甘油兼养不影响转基因藻细胞生长，甘油激酶不表达或表达量降低. 本文探索出了应用基因敲除技术研究三角褐指藻基因功能的研究体系，为今后三角褐指藻功能基因的研究奠定了基础.

Abstract:

Phaeodactylum tricornutum is model organism for marine diatom. It is also one of the most popular energy microalgae. The genome sequence of P. tricornutum had been completed. But, the gene function analysis and gene regulation research of P. tricornutum need further exploration. In this paper, we constructed the glycerol kinase gene knockout vector for P. tricornutum, and successfully converted the vector to wild type P. tricornutum by particle bombardment. After screened, 34 transformations were obtained. The analysis of cell growth showed that extracellular glycerin had no effect on the cell growth of the transformats. Detected by western blot, the results presented that the expression of glycerol kinase were repressed or reduced in transformants. In short, we explored a knockout system for further gene function research in P. tricornutum.