GENETIC-TRANSFORMATION OF MOUSE EMBRYOS BY MICRO-INJECTION OF PURIFIED DNA
GENETIC-TRANSFORMATION OF MOUSE EMBRYOS BY MICRO-INJECTION OF PURIFIED DNA
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DOI：
10.1073/pnas.77.12.7380
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发表时间：
1980-01-01
期刊：
影响因子：
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通讯作者：
RUDDLE, FH
中科院分区：
文献类型：
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作者：
GORDON, JW;SCANGOS, GA;RUDDLE, FH
将由单纯疱疹病毒片段和SV 40病毒DNA组成的重组质粒插入细菌质粒pBR 322中，显微注射到受精小鼠卵母细胞的原核中。将胚胎植入假孕雌性动物的输卵管中，并使其发育至足月。通过Southern印迹技术评价来自新生小鼠的DNA是否存在与注射质粒同源的DNA。在1个系列注射的78只小鼠中，2只小鼠显示出与注射序列的明显同源性，尽管注射序列已重排。2只阳性小鼠的条带强度与新生儿的大多数或所有细胞中存在供体DNA一致。基因显然可以通过直接插入早期胚胎的细胞核而引入小鼠基因组。该技术提供了通过应用重组DNA技术来研究哺乳动物系统中的基因调控和细胞分化问题的机会。
A recombinant plasmid composed of segments of herpes simplex virus and SV 40 viral DNA inserted into the bacterial plasmid pBR322 was microinjected into pronuclei of fertilized mouse oocytes. The embryos were implanted in the oviducts of pseudopregnant females and allowed to develop to term. DNA from newborn mice was evaluated by the Southern blotting technique for the presence of DNA homologous to the injected plasmid. Of 78 mice in 1 series of injections, 2 mice showed clear homology, to the injected sequences, although the injected sequences had been rearranged. Band intensities from the 2 positive mice were consistent with the presence of donor DNA in most or all cells of the newborns. Genes can apparently be introduced into the mouse genome by direct insertion into the nuclei of early embryos. This technique affords the opportunity to study problems of gene regulation and cell differentiation in a mammalian system by application of recombinant DNA technology.