摘要
目的克隆并获得长爪沙鼠载脂蛋白ApoE基因E4外显子的序列。方法根据Genbank中的相关序列，用自行设计的引物，通过常规的RT-PCR方法，从长爪沙鼠肝脏中克隆及测序得到所需要的基因序列。结果长爪沙鼠载脂蛋白ApoE基因E4外显子长约987bp，经比较发现最终测序的结果与大小鼠ApoE基因的同源性达到了73％-74％，与人ApoE基因的同源性达到了62％，与猪、牛ApoE基因的同源性达到了60％以上，与非人灵长类ApoE基因的同源性达到了57％，用DNAstar进行编码氨基酸预测，基因共编码263个氨基酸，已向genbank提交，登录号码EU780067。结论利用基因进化的保守性通过PCR方法可以得到长爪沙鼠载脂蛋白E4基因的序列，本实验为筛选长爪沙鼠高脂血症模型的遗传多样性标记，为培育长爪沙鼠的血脂代谢新品系打下基础。
Objective To clone the apolipoprotein E gene Extron 4 of the Mongolian gerbil. Methods The primers was designed according to the published sequences in GenBank. A RT-PCR method was applied to clone the apolipoprotein E Extron 4 in liver of the Mongolian gerbil. The PCR products was sequenced and blast with related sequences. Results The results of PCR and sequence presented that the sequence of the apolipoprotein E gene Extron 4 in the Mongolian gerbil was a 987 bp sequences. The result of blast showed the identification between Mongolian gerbil and rat or mouse is 73%-74%, between Mongolian gerbil and human is 62%, between Mongolian gerbil and pig or bovine is above 60%, between Mongolian gerbil and no-human primates is 57%. The coding protein is 263 amino acid residuals. Conclusions The RT-PCR products was the sequence of the apolipoprotein E4 gene of Mongolian gerbil.
出处
《实验动物与比较医学》
CAS
2008年第5期289-298,共10页
Laboratory Animal and Comparative Medicine
基金
浙江省自然科学基金(Y3080126)
浙江省医药卫生青年优秀人才基金(2007QN001)
