Kajiwara N, Sugiyama F, Goto Y, Sugiyama Y, Fukamizu A, Uehara S, Sugimura K, Murakami K, Hokao R, Akahori F
Laboratory Animal Research Center, University of Tsukuba, Ibaraki, Japan.
Jikken Dobutsu. 1993 Jul;42(3):463-6. doi: 10.1538/expanim1978.42.3_463.
We used pregnant and pseudopregnant Wistar-Imamichi rats, prepared at a breeding farm, for production of transgenic rats. The donor and recipient rats were transported to our facility for ova manipulation and embryo implantation, respectively. As a foreign gene, 1.9 kb of a hybrid gene was constructed with the rat renin promoter and chloramphenicol acetyltransferase-coding gene. Fertilized oocytes were collected from the donor and the foreign DNA was microinjected into the pronuclei of 539 oocytes by the method of Hochi. The manipulated oocytes were cultured to the 2-cell stage. One hundred seventeen 2-cell embryos were implanted in the recipient rats. Of 67 newborns, 55 rats grew up to be 3-week-old weanlings. Genomic DNA was extracted from the tail of these weanlings and examined by polymerase chain reaction analysis. Six transgenic rats were found to have been generated by the present method. In this way, transgenic rats were produced by an efficient combination of breeding farm utilization and laboratory research.
我们使用在繁殖场培育的怀孕和假孕Wistar-Imamichi大鼠来生产转基因大鼠。供体和受体大鼠分别被运到我们的设施中进行卵子操作和胚胎植入。作为外源基因，用大鼠肾素启动子和氯霉素乙酰转移酶编码基因构建了一个1.9 kb的杂交基因。从供体收集受精卵母细胞，并通过Hochi的方法将外源DNA显微注射到539个卵母细胞的原核中。将操作后的卵母细胞培养到2细胞阶段。将117个2细胞胚胎植入受体大鼠体内。在67只新生大鼠中，55只成长为3周龄的断奶幼鼠。从这些断奶幼鼠的尾巴中提取基因组DNA，并通过聚合酶链反应分析进行检测。发现通过本方法产生了6只转基因大鼠。通过这种方式，通过繁殖场利用和实验室研究的有效结合生产出了转基因大鼠。
