绵羊皮肤成纤维细胞对G418及Blasticidin S抗性的研究

抗生素常被用于对转基因动物、植物及微生物细胞进行筛选。在进行抗生素抗性筛选前,需要摸索合适的抗生素使用浓度,其原因在于:过低的筛选浓度无法抑制非转基因细胞的生长;反之,过高的筛选浓度会导致转基因细胞死亡。为了摸索绵羊皮肤成纤维细胞(sheep skin fibroblasts, SSFs)对G418及Blasticidin S的抗性,本实验以体外培养的SSFs为实验材料,采用不同浓度的上述两种抗生素处理SSFs,采用活细胞计数的方式检测SSFs对上述两种抗生素的抗性。单独使用G418或Blasticidin S导致SSFs全部死亡的最低致死浓度分别为200μg/mL及4μg/mL;当两种抗生素联合使用时,其最低致死浓度为100μg/mL G418+3μg/mL Blasticidin S或150μg/mL G418+2μg/mL Blasticidin S。该实验为采用这两种抗生素筛选转基因SSFs提供了理论基础。

The Study on the Resistance of Sheep Skin Fibroblasts to G418 and Blasticidin S

Antibiotics are often used to screen transgenic animals,plants and microbial cells.Before antibiotic resistance screening,suitable concentration for antibiotics should be explored,which is due to the reason that excessively low screening concentration cannot inhibit the proliferation of non-transgenic cells,on the contrary,excessively high screening concentration will lead to the death of transgenic cells.In order to explore the resistance of sheep skin fibroblasts(SSFs)to G418 and Blasticidin S,SSFs cultured in vitro were used as material in this study.SSFs were treated with different concentrations of above two kinds of antibiotics,and the resistance of SSFs to the above two kinds of antibiotics was determined byliving cell counting method.The results showed that the minimum lethal concentration of SSFs death when used G418 alone or Blasticidin S alone was 200 μg/mL and 4 μg/mL,respectively.When G418 and Blasticidin S were combined,the minimum lethal concentration was 100 μg/mL G418+3 μg/mL Blasticidin S or 150 μg/mL G418+2 μg/mL Blasticidin S.This study could provise a theoretical basis for the screening of transgenic SSFs using these two kinds of antibiotics.