Naito M, Sakurai M, Kuwana T
National Institute of Animal Industry, Tsukuba Norindanchi, Ibaraki, Japan.
J Reprod Fertil. 1998 May;113(1):137-43. doi: 10.1530/jrf.0.1130137.
The expression and fate of exogenous DNA (lacZ gene), introduced into the gonads of chimaeric embryos and chickens that had been produced by transfer of primordial germ cells (PGCs) transfected in vitro, were examined. PGCs obtained from embryonic blood were transfected in vitro by lipofection and transferred to the partially sterilized recipient embryos. Expression of the lacZ gene was observed in the gonads of chimaeric embryos incubated for 3 days after the PGC injection (71.2%, 37/52). Introduction of the lacZ gene into the gonads of chimaeric embryos was confirmed by PCR analysis. The percentage of embryos with gonads positive for the lacZ gene was 95% (38/40) after 3 days of incubation after the PGC injection. The lacZ gene, however, appeared to persist episomally but was gradually lost during embryonic development. After 17 days of incubation after the PGC injection, the lacZ gene was detected in only 14.3% (3/21) of the embryos examined. Although the lacZ gene was detected in the gonads of two hatched chicks (11.1%), it was not detected in the gonads of chimaeric chickens at sexual maturity. Offspring derived from the lipofected PGCs were obtained from the chimaeric chickens at frequencies of 12.1-69.9% in males and 71.6-97.6% in females. The technique developed in the present work could be used to test the expression of exogenous DNA in the gonads of early chicken embryos and should facilitate the production of transgenic chickens.
对通过体外转染原始生殖细胞（PGC）产生的嵌合胚胎和鸡的性腺中引入的外源DNA（lacZ基因）的表达和命运进行了检测。从胚胎血液中获得的PGC在体外通过脂质体转染，然后转移到部分绝育的受体胚胎中。在PGC注射后培养3天的嵌合胚胎性腺中观察到lacZ基因的表达（71.2%，37/52）。通过PCR分析证实了lacZ基因已导入嵌合胚胎的性腺中。PGC注射后培养3天，性腺中lacZ基因呈阳性的胚胎百分比为95%（38/40）。然而，lacZ基因似乎以附加体形式持续存在，但在胚胎发育过程中逐渐丢失。PGC注射后培养17天，在所检测的胚胎中仅14.3%（3/21）检测到lacZ基因。虽然在两只孵化出的雏鸡的性腺中检测到了lacZ基因（11.1%），但在性成熟的嵌合鸡的性腺中未检测到。来自脂质体转染PGC的后代在雄性嵌合鸡中的获得频率为12.1 - 69.9%，在雌性嵌合鸡中的获得频率为71.6 - 97.6%。本研究中开发的技术可用于检测外源DNA在早期鸡胚胎性腺中的表达，并应有助于转基因鸡的生产。
