慢病毒载体介导的转人ahsp基因的β^(654)地中海贫血小鼠的制备  

Generation of Human ahsp Transgenic β^(654)-Thalassemia Mice Mediated by Lentiviral Vector

作　　者：王宝斌[1] 方彧聃[1] 郭歆冰[1] 任兆瑞[1] 张敬之[1] 

机构地区：[1]上海交通大学医学遗传研究所,卫生部医学胚胎分子生物学重点实验室暨上海市胚胎与分子生物学重点实验室,上海200040

出　　处：《生物技术通讯》2008年第6期833-835,共3页Letters in Biotechnology

基　　金：国家自然科学基金(30571777);国家重点基础研究发展计划(2004CB518806);国家高技术研究发展计划(2007AA021206)

摘　　要：目的:经慢病毒载体介导,制备转人α血红蛋白稳定蛋白基因(ahsp)的β654地中海贫血小鼠。方法:用巢式PCR从人血DNA中获得人ahsp基因,构建含有人ahsp基因的慢病毒载体,制备假病毒,通过卵周隙显微注射手段将其导入β654地贫小鼠的受精卵,经移植至假孕母鼠输卵管,最终孕育出转人ahsp基因的β654地贫小鼠;分析小鼠体内外源ahsp基因的表达情况及其遗传稳定性。结果:共获得了8只人ahsp阳性小鼠,转基因阳性率为32%(8/25),其中3只同时具有β654突变基因;人ahsp基因在小鼠体内的表达水平维持在小鼠自身ahsp表达量的1%左右,且可稳定遗传至子代。结论:制备了转人ahsp基因小鼠,并可遗传至子代,为在个体水平上研究α血红蛋白稳定蛋白与β地贫之间的关系提供了工具。Objective： To obtain human α-hemoglobin stabilizing protein（ahsp） transgenic β654-thalassemia mice. Methods： Human ahsp gene with its promoter was amplified by nested-PCR from human blood geoomic DNA, then it was cloned into a lentiviral vector to construct its pseudo virus. The human ahsp transgenic mice were subsequently generated by infecting the fertilized oocytes of β654-thalassemia mice with pseudo virus. The transcription of the human ahsp gene and its genetic satiability were investigated. Results： We obtained 8 human ahsp transgenic mice, among which, 3 carried β654 gene. mRNA quantity of human ahsp accounted for approximately 1% of that of mouse ahsp. The transgene could be in- herited to the next generation. Conclusion： Human ahsp transgenic mice can be obtained by the method described herein, and they can be useful for studying the relationship between α-hemoglobin stabilizing protein and β654-thalassemia.

关 键 词：α血红蛋白稳定蛋白基因 β654地中海贫血 慢病毒载体 转基因小鼠 

分 类 号：Q789[生物学—分子生物学]