转TNF-α基因鱼腥藻7120质粒稳定性研究

Study on Plasmid Stability of Recombinant Anabaena sp.PCC7120

为了实现转基因鱼腥藻培养生产TNF的目的 ,研究了转基因鱼腥藻的稳定性。影印法证实转TNF -α基因鱼腥藻 712 0能保持质粒分配稳定性。比较无选择压力下连续传代的转基因鱼腥藻 712 0在不同培养基中的生长和外源基因表达 ,证实没有发生质粒部分缺失 ,但转基因鱼腥藻在无选择压力下会降低重组质粒拷贝数。在培养过程中 ,种子培养基含有的新霉素可以保持生产过程质粒稳定 ,这可以大大减少新霉素用量。

The plasmid stability of recombinant Anabaena sp.PCC7120 was studied.The experiment proved that recombinant Anabaena sp.PCC7120 could remain segregative stability of recombinant plasmid pDC-TNF.Through comparing the growth and exogenous gene expression of recombinant Anabaena sp.PCC7120 that existed from generation to generation without neomycin and cultured in different medium,it is also proved that the structural stability of recombinant plasmid pDC-TNF was high,but the copy number of recombinant plasmid pDC-TNF would reduce if recombinant Anabaena sp.PCC7120 was cultured in medium without neomycin.The neomycin in the seed medium could hold the stability of pDC-TNF during culture process,so the quantity of neomycin used in fermentation culture could be reduced much.