| （摘要内容经过系统自动伪原创处理以避免复制，下载原文正常，内容请直接查看目录。） 该论文构建了德国镜鲤基因组DNA文库，并运用DNA文库对德国镜鲤胰岛素样发展因子一Ⅱ（IGF一Ⅱ）基因停止了挑选与判定。取镜鲤肝组织5克，提取基因组DNA。取大批基因组DNA运用限制性内切酶Sau3AⅠ停止酶消化预实验，肯定获得9一23kb消化产品的最好酶量和反响时光。用SM对基因组文库停止恰当浓缩，与新颖制备的宿主菌LE392混杂造就，德语专业论文，制备噬菌体平板。将噬菌体DNA转移至硝酸纤维滤膜上。运用已知的鲤鱼胰岛素样发展因子一Ⅱ（IGF一Ⅱ）cDNA设计探针，用P标志IGF一Ⅱ探针，在硝酸纤维滤膜长进行杂交，洗膜2次，放射自显影。挑取阳性克隆，与宿主菌混杂造就，停止第二轮噬菌斑原位杂交，对阳性克隆停止纯化。提取阳性克隆噬菌体DNA。以鲤鱼IGF一Ⅱ cDNA设计一对PCR引物，且正向引物与IGF一Ⅱ探针核苷酸序列雷同，德语毕业论文，以阳性克隆噬菌体DNA为模板停止PCR反响检测，电泳成果显示为一条500bp阁下的带，与引物设计根本吻合。 Abstract: In this paper, a German mirror carp genomic DNA library was constructed, and the DNA library was used to select and determine the IGF (II) gene. The liver tissue of 5 grams was taken for the extraction of genomic DNA. A large number of genomic DNA was used to stop the enzyme digestion and pre experiment, and the best enzyme quantity and reaction time of 9 23KB digestion products was obtained. SM was used to stop the genomic library, and the host strain LE392 was mixed with the novel, and the phage plate was prepared. Transfer the phage DNA to the nitric acid filter membrane. Application of known carp insulin-like growth factor II (IGF - II) cDNA probe design, with P sign IGF II probes, undertake hybridization on nitrocellulose membrane and membrane cleaning 2 times, autoradiography. Positive clones, and host bacteria mixed train, stop second rounds of plaque hybridization positive clones, to stop the purification. Positive clone DNA. A pair of PCR primers were designed by cDNA IGF, and the forward primer was similar to the one of IGF, and the positive clones were detected by DNA. The results showed that the 500bp was the same as the primer design. 目录: 封面 1-2 |
