作　　者： ; ; ; ; ;

机构地区： 汕头大学医学院炎症与免疫性疾病研究所

出　　处： 《医学研究生学报》 2008年第5期458-460,I0001,共4页

摘　　要： 目的:克隆小鼠可溶性ST2基因并构建其真核表达载体。方法:从小鼠肥大细胞株P815中提取总RNA,逆转录为cDNA,用热启动PCR技术,扩增小鼠可溶性ST2基因全长编码区,经双酶切后,克隆入pcDNA3.1载体中,构建真核表达载体pcDNA3.1-mST2,然后进行酶切鉴定和序列分析。结果:小鼠可溶性ST2基因的PCR产物和重组载体经凝胶电泳和酶切鉴定、测序分析证实,其序列与GenBank中数据一致。小鼠可溶性ST2基因的全长编码序列为1 011 bp,编码336个氨基酸。结论:用热启动PCR技术能方便、准确地获得目的基因片段。成功地克隆小鼠可溶性ST2基因并构建了其真核表达载体,为进一步进行小鼠可溶性ST2蛋白的表达和功能研究奠定了基础。 Objective： To clone the mouse soluble ST2 gene and construct its eukaryotic expression vector. Methods： Total RNA was prepared from P815 cells, and the cDNA fragment encoding the soluble ST2 gene was amplified by RT-PCR using specific primers and then cloned into the pcDNA3.1 vector. The recombinant eukaryotic expression vector pcDNA3.1-mST2 was constructed and was transferred into the host E. coli strain TOP10 for identification. And the inserted soluble ST2 gene was sequenced. Results： Sequence analysis indicated that the full length of the mouse soluble ST2 gene consisted of 1011 bp, encoded 336 amino acids and had a high nucleotide homology with that reported previously. Con- clusion： We was successfully cloned the mouse soluble ST2 gene and constructed its eukaryotic expression vector, which has paved the way for further studies on the expression and function of the soluble ST2 in mice.

关 键 词： 小鼠可溶性 热启动 基因克隆

领　　域： [生物学]