家族性肌萎缩侧索硬化症转基因鼠的繁殖和鉴定

目的 建立家族性肌萎缩侧索硬化症（FALS）转基因模型鼠的繁育方法，并对其子代鼠进行基因鉴定。方法 （1）以6只B6SJLSOD1G93～＋半合子雄鼠与6只B6SJLFIJ＋／＋雌鼠（1：1）交配；（2）由鼠尾血提取基因组DNA，PCR扩增hmSOD1基因的片段，电泳后观察结果；（3）对PCR产物进行纯化测序，并通过BLAST验证。结果 6对种鼠交配产鼠仔53只，存活率为98％（52／53）；G93AhmSODl阳性鼠约占44．2％（23／52）；PCR扩增产物分别为内对照（IL-2）：324bp；Tg（hmSOD1）：236bp；测序证实PCR产物的基因序列和hmSODl基因片段中的序列一致．并存在G93A突变。结论 B6SJL-Tg（SOD1-G93A）1Gur／J雄鼠与B6SJLFI／J雌鼠配种能成功繁育出Tg（hmSOD1）阳性的ALS半合子子代鼠：本实验的PCR法能准确鉴定hmSOD1基因阳性鼠，并证实该转人的基因按近似孟德尔方式遗传，为ALS实验研究奠定了基础。

Establishment of transgenic mouse model of familial amyotrophic lateral sclerosis and identification of the filial generation

Objective To establish transgenic mouse models of familial amyotrophic lateral sclerosis(FALS) and identify the genotype of the first filial generation.Methods Six male B6SJL SOD1G93A/ hemizygote mice were mated with 6 female B6SJLF1/J / mice to produce the filial generation.The genomic DNA was extracted from the tail vein blood of the first filial generation mice and PCR was performed to amplify the hmSOD1 gene fragment.The genotype of the mice was determined by electrophoresis,and the PCR product was purified for further gene sequence analysis and detection of mutation loci.Results Fifty-three progeny mice were born and the survival rate before ALS onset was 98%(52/53),and among the survived mice,the positivity rate for hmSOD1 gene was 44.2 %(23/52).Electrophoresis result showed that the PCR product of 236 bp was consistent with the hmSOD1 gene fragment,and the sequence of the PCR product was identical with hmSOD1 gene sequence of G93A mutant.Conclusion Transgenic mouse models of ALS can be established in the first filial generation of male B6SJL SOD1G93A/ mice mated with female B6SJLF1/J / .PCR technique can precisely identify the genotype of the filial generation.