SOD1-G93A转基因小鼠的鉴定方法比较

Comparison of SOD1-G93A transgenic mice identification methods

目的建立一种简便、节省和准确的肌萎缩侧索硬化(ALS)hSOD1-G93A转基因小鼠DNA提取及基因鉴定方法。方法以B6SJL-BTg(SOD1-G93A)1Gur/J半合子雄鼠与B6SJLF1雌鼠交配产仔,剪尾尖约1mm,提取组织DNA,PCR扩增hSOD1基因片断,电泳后观察结果并对PCR产物进行纯化、测序及BLAST验证。同时留取鼠尾血50～100μl,提取血DNA并PCR扩增作为对照,比较两者之间的符合率。结果hSOD1-G93A阳性鼠约占54%,基本符合孟德尔遗传规律。鼠尾组织及血DNA鉴定结果符合率为100%。结论鼠尾尖组织DNA提取及鉴定是一种简便易行、节省经费并且准确可靠的方法。

Objective To estabolish a convenient, sparing and accurate DNA extraction and gene identification method of hSOD1-G93A transgenic mice. Methods B6SJL-Tg（SODI-G93A） 1Gur/J hemizygote male mice were mated with B6SJLF1 female mice. About lmm of the tip of mice tail was cut and DNA was extracted. hSOD1 gene segment was PCR amplificated. The genotype of the mice was determined by electrophoresis, and PCR product was purified and sequenced to verify the mutantion. At the same time, DNA of 50 - 100μl blood from each mice tail was used as positive control to test the accuracy of tissue DNA. Results hSODI-G93A positive mice were 54% , basically consistent with Mendelian inheritance rule. The results of tail and blood DNA identification were completely coincident. Conclusion DNA extraction and gene identification from tail tip tissue was a convenient, cheao and accurate method.