应用PCR测定Cx43转基因小鼠的基因型

GENOTYPING CONNEXIN 43 TRANSGENIC MICE WITH PCR

目的 :应用聚合酶链反应 (PCR)技术 ,检测 Cx4 3(connexin 4 3)转基因小鼠的基因型。方法 :采用 CMV4 3转基因小鼠和Cx4 3剔除小鼠分别与野生型小鼠杂交繁殖 ,切取子代小鼠尾巴制备 DNA,应用 PCR技术扩增 DNA,观察目的基因的阳性率。结果 :CMV4 3转基因小鼠子代 6 6个 ,dhfr PCR检测阳性率为 31.8%。 Cx4 3基因剔除小鼠子代 2 1个 ,分别与 Cx4 3野生型引物和 Cx4 3剔除引物进行 PCR,两者均阳性 (+ / + ) 12个 ,百分率为 5 7.1% ,前者阳性而后者阴性 (杂合子 + / - ) 7个 ,百分率为 33.3% ,两者均阴性 (纯合子 - / - ) 2个 ,百分率为 9.6 %。野生型小鼠经 Cx4 3野生型引物 PCR检测结果均为阳性。结论 :PCR技术用于 Cx4 3转基因小鼠基因型的检测敏感度高 ,特异性强 ,易操作 ,可用作 Cx4 3转基因小鼠心血管畸形模型研究的初选方法。

Objective:To genotype transgenic mice with polymerase chain reaction (PCR) technique. Methods:The tails of the CMV43 transgenic and connexin 43 (Cx43 ) knockout mice were cut to pick up DNA. Then the DNA was amplified by PCR and the positive ratio of the target gene was analyzed.Result:The CMV43 transgenic mice were 66, which the positive ratio by dhfr PCR was 31.8 per cent. The Cx43 knockout mice were 21, which were amplified by PCR with the primers of wild type and Cx43 knockout, respectively. Twelve was all positive( +/ +) with 57.1 percent, 7 heterozygous( +/ -) with 33.3 percent, 2 homozygous( -/ -) with 9.6 percent. Wild type mice were all positive.Conclusion:PCR technique is a highly sensitive, specific, and easier to genotype Connexin 43 transgenic mice. It may be the primary method to study Connexin 43 transgenic mice for cardiovascular malformation model.