猪胚胎成纤维细胞αGal表达的鉴定  

Primary identification of the expression of αGal epitope in porcine embryonic fibroblast

作　　者：王坤[1] 庄国华[1] 张同琳[1] 林爱星[2] 周孝思[1] 

机构地区：[1]北京大学第三医院器官移植中心,100083 [2]中国农业大学农业生物技术国家重点实验室

出　　处：《中华器官移植杂志》2005年第9期521-523,共3页Chinese Journal of Organ Transplantation

摘　　要：目的确定猪胚胎成纤维细胞是否合成α1,3半乳糖苷,为下一步进行基因工程改造的结果提供鉴定方法。方法采用逆转录聚合酶链反应(RT-PCR)和Western免疫印迹法分别在转录和翻译水平确定猪胚胎成纤维细胞是否合成α1,3半乳糖苷。结果通过RT-PCR扩增,发现α1,3半乳糖苷转移酶基因在原代和传代培养的猪胚胎成纤维细胞内转录,Western免疫印迹证实,在原代和传代培养的猪胚胎成纤维细胞能合成含有α1,3半乳糖苷残基的糖蛋白。结论猪胚胎成纤维细胞可合成α1,3半乳糖苷;可采用PCR扩增和Western免疫印迹方法对经过基因工程改造的猪胚胎成纤维细胞是否表达α1,3半乳糖苷转移酶进行鉴定。Objective To study the synthesis of α1,3galactosyltransferase gene in porcine embryonic fibroblast. Methods The transcription and translation of α1,3galactosyltransferase gene were identified in porcine embryonic fibroblast by RT-PCR and Western blot. Results It was identified that there was the expression of α1,3 galactosyltransferase gene in the cultured porcine embryonic fibroblasts. Conclusion α1,3 galactosyltransferase gene can be synthesized in porcine embryonic fibroblast. RT-PCR and Western blot can be applied to identify the expression of α1,3 galactosyltransferase gene in the porcine embryonic fibroblast.

关 键 词：猪 胚胎 成纤维细胞 半乳糖转移酶类 鉴定方法 猪胚胎 Western免疫印迹方法 逆转录聚合酶链反应 RT-PCR扩增 GAL 

分 类 号：R346[医药卫生—基础医学]