论文部分内容阅读
目的人反义ｉｇｆⅡ基因真核表达载体的构建．方法ＰＣ／ｇｅｎｅ软件分析并设计人ＩＧＦⅡｃＤＮＡＢ区域，化学合成并得到人ＩＧＦⅡｃＤＮＡ克隆于原核载体ｐＧＥＭ７Ｚｆ（＋），将之反向连接到逆转病毒表达载体ｐｃＤＮＡ３．经定向设计，将ＩＧＦⅡ反义基因插入真核载体，ｐｃＤＮＡ３中．结果经ＤＮＡ双链测序证明，克隆的人ＩＧＦⅡｃＤＮＡ序列与设计完全一致，构建所得反义ＩＧＦⅡ真核载体ｐＩＧＦⅡＡｓ，经Ｄｏｔｂｌｏｔ，ＰＣＲ鉴定结果正确．结论为小片断的基因合成与克隆提供了简捷可靠的方法，得到了反义ｉｇｆⅡ基因真核表达载体，为肝癌的反义基因治疗提供了基础
Construction of eukaryotic expression vector of target human antisense igf-Ⅱ gene. Methods PC / gene software was used to analyze and design the human IGF-ⅡcDNAB region. The human IGF-Ⅱ cDNA was chemically synthesized and cloned into the prokaryotic vector pGEM7Zf (+), which was reversely ligated to the retroviral expression vector pcDNA3. The directional design, the IGF Ⅱ antisense gene into eukaryotic vector, pcDNA3. Results DNA double-stranded sequencing proved that cloned human IGF-Ⅱ cDNA sequence and design exactly the same, construct the resulting antisense IGF Ⅱ eukaryotic vector pIGF Ⅱ As by Dotblot, PCR identification results correct. Conclusion The method provided a simple and reliable method for gene synthesis and cloning of small fragments. The eukaryotic expression vector of antisense igf Ⅱ gene was obtained, which provided the basis for antisense gene therapy of liver cancer