人反义igf-Ⅱ基因真核表达载体pIGF-ⅡAs的构建  

Construction of pIGFⅡAs:a eukaryotic expression vector for antisense human insulinlike growth factor Ⅱ gene*

作　　者：张鸣青[1,2] 杨冬华[1,2] 毛积芳 顾红[1,2] 徐重 

机构地区：[1]第一军医大学珠江医院消化内科 [2]第二军医大学生物化学教研室

出　　处：《华人消化杂志》1998年第9期776-779,共4页

基　　金：广东省科委自然科学基金

摘　　要：目的人反义ｉｇｆⅡ基因真核表达载体的构建．方法ＰＣ／ｇｅｎｅ软件分析并设计人ＩＧＦⅡｃＤＮＡＢ区域，化学合成并得到人ＩＧＦⅡｃＤＮＡ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ＩＧＦⅡ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ＩＧＦⅡｃＤＮＡ序列与设计完全一致，构建所得反义ＩＧＦⅡ真核载体ｐＩＧＦⅡＡｓ，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ｉｇｆⅡ基因真核表达载体。IM Abstract:To construct a eukaryotic expression vector for antisense human insulin like growth factor Ⅱ (IGFⅡ) gene.METHODS A cDNA coding B domain of the human IGFⅡ was synthesized and cloned into pGEM7Zf(+) vector. The IGFⅡ fragment from restriction endonuclease digestion was further inserted into eukaryotic vector pcDNA3 in transdirection.RESULTS The sequence of cloned IGFⅡ cDNA was proved to be the same as the designed. The constructed antisense IGFⅡ pcDNA3 was identified to be correct by PCR and dot blotting hybridization.CONCLUSION A reliable and forthright method for chemical synthesis of small fragments gene and cloning was provided. A way for construction of pIGFⅡAsA eukaryotic expression vector for antisense human IGFⅡ gene was set up. This study is one of the basic work for IGFⅡ antisense gene therapy for hepatocellular carcinoma.

关 键 词：真核细胞 遗传学 基因结构 免疫球蛋白 

分 类 号：R394[医药卫生—医学遗传学]