【摘要】 目的 :构建具足够库容量的小鼠牙胚 c DNA文库。方法 :先提取小鼠牙胚 m RNA并反转录成c DNA,经 L D PCR扩增后 ,将双链 c DNA克隆到 λTriple X中 ,包装噬菌体 ,以 XL- l blue为受菌体滴定、扩增文库 ,用已知的牙胚基质蛋白基因引物作 PCR鉴定文库。结果 :5 0 0 μl文库的库容量为 1.3× 10 6 pfu,重组率为 72 % ,从文库中克隆到已知的 5种牙胚基蛋白基因。结论 :构建的文库具较好的库容量和重组率及较大的插入片段。更多还原

【Abstract】 Objective:To establish a cDNA library of mouse dental germ.Method:Extraction of mRNA of mouse dental germ and synthesis of cDNA were conducted with reverse DNA polymerase.Then the ds cDNA was ligated to TripleX and the recombinant bacteriophage was packaged.The cDNA library was titered and amplified with XL l blue as receptor bacterium and identified with 5 pairs of primers of genes of mouse dental germ matrix proteins by PCR.Result:The titer of the unamplified library was 1.3×10 6 pfu,and the percentage of recombinant clones was 72%.And all the five known genes were contained in the library,the size of the fragment was 670～1900 bp.Conclusion:The established cDNA library has high titer ,recombinant prcentage and large insert fragments of genes.更多还原