Construction Of CDNA Library And IL-18 Gene Cloning And Expression Of Beijing Fatty Chicken

The cDNA library Construction of Beijing Fatty Chicken has great significance hot only for its protection of genetic resources but also for the reaseach of gene function.In this study,the total RNA were extracted from the liver tissure of Beijing Fatty Chicken, and the liver tissure cDNA library of Beijing Fatty Chicken had been constructed by using SMARTTM technique.The results show that the titer of unamplified cDNA library is 2.01×106pfu·mL-1, the titer of amplified library is 1.17×1010pfu·mL-1, the rate of recombinant is above 93.8%, the average size of the fragments is 1.0 kb.This study is not only important for the structure and function research on Chicken genes,but also important for the protection of genetic resources for Beijing Fatty Chicken.This study successfully established the target cell line of Duolang Sheep through direct adherent culture metheods and also successfully conducted the various kinds of experiments on the biological characteristics of detection.The results show that the quality of the cell line met the quality requirements of the ATCC (American Type Culture Collection), indicate that the cells were well target cells for gene expression.In this study , we also constructed Beijing Fatty Chicken's Prokaryotic expression vector pGEX-4T-1-IL-18 by adopting BamH I and Xho I method of double restriction enzyme and which are also expressed in BL21 bacteria, we also constructed Beijing Fatty Chicken's eukaryotic expression vector pEGFP-N3-IL-18 by adopting Sal I and BamH I method of double restriction enzyme and expression in target cells.