Xie H X
Cell Struct Funct. 1983 Dec;8(4):315-25. doi: 10.1247/csf.8.315.
The efficiency of an exogenous gene's expression was compared after its transfection and injection into various mouse cells to systematically evaluate these two gene transfer techniques. Special attention was paid to the period of transient expression. The gene used was a derivative of chicken delta-crystallin gene with the 5' end region replaced by a promoter base sequence of a retrovirus. Nuclear injection was more efficient than transfection in several respects: it was roughly one thousand times more efficient in producing gene-expressing cells than the transfection technique; it produced positive cells in every challenged cell line in contrast to the results of some unsuccessful trials found with transfection; and the maximum expression of the exogenous gene in a gene-transferred cell was much higher after injection than after transfection. With the transfection technique, use of a DNA-calcium phosphate coprecipitate was slightly more efficient than the use of DEAE-dextran. The stability of gene expression in transfected and nuclear-injected cells differed greatly: Expression of the exogenous gene in transfected cells was transmitted to 92% of the daughter cells per division, whereas its expression in injected cells was transmitted to only 32% of the daughter cells. This great difference in stability probably reflects different states of the major fraction of the exogenous gene: integration into chromosomes in transfected cells versus extrachromosomal localization in injected cells.
将外源基因转染并注射到各种小鼠细胞后，比较其表达效率，以系统评估这两种基因转移技术。特别关注瞬时表达期。所用基因是鸡δ-晶体蛋白基因的衍生物，其5'端区域被逆转录病毒的启动子碱基序列取代。核注射在几个方面比转染更有效：在产生基因表达细胞方面，它比转染技术效率高约一千倍；与转染的一些不成功试验结果相反，它在每个受挑战的细胞系中都产生了阳性细胞；基因转移细胞中外源基因的最大表达在注射后比转染后高得多。使用转染技术时，DNA-磷酸钙共沉淀物的效率略高于DEAE-葡聚糖。转染细胞和核注射细胞中基因表达的稳定性差异很大：转染细胞中外源基因的表达在每次分裂时传递给92%的子细胞，而其在注射细胞中的表达仅传递给32%的子细胞。这种稳定性的巨大差异可能反映了外源基因主要部分的不同状态：转染细胞中整合到染色体中，而注射细胞中位于染色体外。
