CAJ | 학술논문

目的：繁殖和鉴定携带有外源LRP16基因的转基因小鼠，为体内研究LRP16的生物学功能提供动物模型。方法首先将构建好的LRP16转基因重组质粒转染到293T细胞，鉴定该质粒是否可以有效表达目的蛋白，随后利用原核显微注射方法将线性化的重组质粒注射入品系为FVB的小鼠受精卵中，获得F0代小鼠。应用PCR方法对F0代小鼠进行鉴定，并将阳性F0代小鼠继续繁殖。结果 LRP16转基因重组质粒在293T细胞中可以有效表达，经PCR鉴定获得3只阳性F0代小鼠，阳性率为6.23%，选取其中1只整合有LRP16基因的F0代雄鼠，通过其与野生型FVB小鼠杂交获得7只F1代子鼠，其中4只为阳性鼠，阳性率为57.14%。从F1代中再次选取其中1只阳性雄鼠与野生型小鼠杂交繁殖获得5只F2代子鼠，PCR鉴定结果显示2只为阳性，阳性率为40%。结论 LRP16基因成功整合到小鼠基因组中，并且能够稳定遗传，为后续在动物整体水平研究LRP16的功能奠定了基础。
목적：번식화감정휴대유외원LRP16기인적전기인소서，위체내연구LRP16적생물학공능제공동물모형。방법수선장구건호적LRP16전기인중조질립전염도293T세포，감정해질립시부가이유효표체목적단백，수후이용원핵현미주사방법장선성화적중조질립주사입품계위FVB적소서수정란중，획득F0대소서。응용PCR방법대F0대소서진행감정，병장양성F0대소서계속번식。결과 LRP16전기인중조질립재293T세포중가이유효표체，경PCR감정획득3지양성F0대소서，양성솔위6.23%，선취기중1지정합유LRP16기인적F0대웅서，통과기여야생형FVB소서잡교획득7지F1대자서，기중4지위양성서，양성솔위57.14%。종F1대중재차선취기중1지양성웅서여야생형소서잡교번식획득5지F2대자서，PCR감정결과현시2지위양성，양성솔위40%。결론 LRP16기인성공정합도소서기인조중，병차능구은정유전，위후속재동물정체수평연구LRP16적공능전정료기출。
Objective To establish the animal model for studying the in vivo biological function of LRP16 by reproducing and identifying LRP16 transgenic mice. Methods After the constructed LRP16 transgenic recombinant plasmid was transfected into 293T cells, whether it can effectively express the target protein was identified. The linear recombinant plasmid was injected into the fertilized ovum of FVB mice to produce F0 mice. The positive F0 mice were identified by PCR and reproduced. Results The constructed recombinant LRP16 plasmid was effectively expressed in 293T cells. Three positive F0 mice were identified by PCR with a positive rate of 6.23%. One male F0 mouse with the integrated LRP16 gene was hybridized with wild FVB mice to reproduce 7 F1 mice, 4 out of which were positive with a positive rate of 57.14%. One male F1 mouse with the integrated LRP16 gene was hybridized with wild FVB mice to reproduce 5 F2 mice, 2 out of which were positive identified by PCR with a positive rate of 40%. Conclusion LRP16 gene is successfully integrated into the mouse genome and can maintain its inheritance, thus laying a foundation for further studying the LRP16 function in animals.