中文摘要：

藻蓝蛋白（Phycocyanin）具有抗氧化、消炎、抗癌、增强免疫等生理活性和荧光特性.本研究从钝顶节旋藻（Arthrospira platensis）中克隆藻蓝蛋白基因,构建重组藻蓝蛋白并在聚球藻7942（Synechococcus sp.PCC 7942）中表达.利用PCR技术克隆钝顶节旋藻藻蓝蛋白α亚基（cpc A）与β亚基（cpc B）基因,在cpc A的5’端及3’端加入编码6×His标签基因,克隆Hiscpc A基因,并构建同源重组表达载体p U BCA3D-1.通过自然转化法将p UBCA3D-1转入聚球藻中,经筛选鉴定获得了转基因藻株.PCR验证结果表明,目的基因整合到聚球藻基因组中的概率为86.6%.转基因聚球藻在无抗生素筛选条件下连续培养2个月,仍能稳定表达重组Cpc A.以上结果表明,重组钝顶节旋藻藻蓝蛋白在聚球藻中稳定表达;研究结果为节旋藻cpc A基因改造、重组Cpc A进一步分离纯化以及重组藻蓝蛋白的生产奠定了基础.

英文摘要：

Phycocyanin consists of α subunit and β subunit with fl uorescence characteristics and physiological characteristics of antioxidant, anti-cancer and increasing immune activity. This research cloned phycocyanin gene from Arthrospira platensis, and constructed the recombinant phycocyanin gene in Synechococcus sp. PCC 7942. The genes of phycocyanin α subunit（cpc A） and β subunit（cpc B） from Arthrospira platensis were cloned by using PCR technology. By adding the 6 × His Tag genes in the 5＇ end and 3＇ end of cpc A, the recombinant 6 × Hiscpc A gene was constructed. Finally the homologous recombination expression vector p UBCA3D-1 was obtained. By using the natural transformation method, the expression vector wa s transformed into Synechococcus, and transgenic algae strains obtained by screening conditions. The PCR identification results showed the rate of target genes integrated into the genome as 86.6%. Under conditions of continuous cultivation without antibiotic selection for two months, transgenic algae strains could still stably express the recombinant phycocyanin. These results demonstrated that the recombinant phycocyanin was successfully expressed in Synechococcus. The results of the study laid a foundation for Arthrospira cpc A genetic modification and recombinant Cpc A purification, playing an important role in production and purification of recombinant phycocyanin.