小鼠α1,3-半乳糖基转移酶基因的克隆及测序  

Cloning and Sequencing of the Murine α1,3-galactosyltransferase Gene

作　　者：杨明浩 郭军 

机构地区：[1]昆明总医院急诊ICU,昆明650032

出　　处：《昆明医学院学报》2006年第3期19-22,共4页Journal of Kunming Medical College

摘　　要：目的克隆α1,3-半乳糖基转移酶的序列,为进一步利用其构建真核表达载体,对肿瘤进行基因治疗奠定基础.方法从BALB/c小鼠腹腔巨噬细胞中提取总RNA,行RT-PCR扩增出编码α1,3-半乳糖基转移酶的全长cDNA,并克隆入pUCm-T测序载体.经DNA测序证实序列.结果经RT-PCR扩增出大小约为1.2kb的基因片段,成功克隆入pUCm-T载体.结论成功地克隆α1,3-半乳糖基转移酶基因序列,为进一步肿瘤基因治疗奠定了基础.Objective To clone α 1,3 - galaetosyltransferase gene and provide the premise for construction of its recombinant eukaryotie plasmid in gene therapy of cancer. Methods Total RNA was isolated from the peritoneal maerophages of BALB/c mice. Subsequently, the full length eDNA of α1,3 - galaetosyltransferase gene was amplified by RT - PCR and cloned into pUCm - T vector confirmed by DNA sequencing. Results A 1.2 kb fragment of α 1,3 - galaetosyltransferase gene was successfully amplified by RT - PCR and cloned into pUCm -T vector. DNA sequencing suggested that the α1,3 -galactosyltransferase gene was 1 221 bp in length, possessing correct read frame and nueleotide sequence. Conclusions The α 1,3 - galaetosyltransferase gene was successfully cloned and this has laid a solid foundation for tumor gene therapy.

关 键 词：Α1 3-半乳糖基转移酶 Galα(1 3)Gal RT-PCR 克隆 

分 类 号：R735.2[医药卫生—肿瘤]