植酸酶基因在海水小球藻细胞内的表达

为建立海水小球藻外源基因转化系统,使外源植酸酶基因在海水小球藻细胞内获得稳定表达,利用电激法将植酸酶基因与海水小球藻细胞共转化;利用含G418的抗性平板筛选抗性藻株,并对所获得的抗性藻株依次进行PCR检测、Southern杂交分析和植酸酶活性与性质的检测.经过转化处理的微藻细胞在含G418的抗性平板上培养3周后,每个平板有10~30个抗性藻株长出;在这些抗性藻株中,PCR检测的阳性率为40.16%;Southern杂交分析和植酸酶活性检测证明,植酸酶外源基因已经在小球藻细胞中获得正确表达.植酸酶外源基因在海水小球藻细胞内被成功表达.

Expression of phytase gene in the cells of marine unicellular microalga chlorella Sp. MACC/C95

To set up a transformation system for the expression of foreign phytase gene in a kind of marine unicellular green microalga(Chlorella sp.MACC/C95),the microalgal cells were transformed with a plasmid containing the phytase gene under the control of CaMV35S promoters and the neomycin phosphotransferase(nptⅡ) gene as a selectable gene by the method of electroporation,and the strains of transformation were selected out by the plate of G418,and then all of transformation strains were checked by the PCR and Southern blotting analysis.Results indicated that about 10-30 G418-resistant strains were found on each plate of G418 after being cultured for three weeks;40.16% of the G418-resistant strains were transformed,and the phytase gene had been integrated into the Chlorella sp.Genome.The expression of phytase protein in transformants was confirmed by the analysis of phytase activity,which showed that the foreign phytase genes had been successfully expressed in the cells of Chlorella sp.MACC/C95.