생쥐 초기배의 pSV - LacZ Reporter 유전자 발현에 관한 연구 = Expression of pSV - LacZ Reporter Gene in Preimplantation Mouse Embryos

다국어 초록 (Multilingual Abstract)

DNA containing the SV40 as an early promoter and enhancer coupled to the structural gene for Escherichia coli β-galactosidase (LacZ) was microinjected into the pronuclei of one-cell mouse embryos. A qualitative histochemical assay, with 5-bromo-4-chloro-3-indoly β-D-gatactoside (X-Gal) as a substrate, was used to detect expression of the LacZ gene at several preimplantation stages. The objectives of this study were to determine in vitro developmental rates for low and high DNA concentrations and expression of the LacZ reporter gene. After microinjection with a lower concentration DNA of 2 ㎍/㎖, 46.0% of the fertilized eggs survived to become the two-cell stage embryos, while 25.1 % and 30.4% developed to floe morula, and blastocyst stage, respectively. At the higher DNA concentration of 319 ㎍/㎖, 51.5% survived into two-cell stage embryos. However, none developed to 8-cell stage embryos. Subsequently, on microirtjecting the higher DNA concentration, LacZ was expressed in 22.5% of the two-cell stage embryos. Also, on microinjecting the lower DNA concentration, LacZ was expressed in 37.2% and 11.5% of the morula and blastocyst stages respectively. The results indicate that developmental rates of embryos decreased as the DNA concentration was increased. This indicates that expression of the fusion gene with SV40 promoter depended on the higher concentration of DNA injected. Altogether, these results indicated that the LacZ reporter gene could be a cell lineage capable of being used as a marker to study cells in mammalian development. Thus, the DNA microinjection technique should be useful for the production of transgenic mice.