白血病分化相关基因dif14的原核表达

Expression of leukemia cell differentiation related gene dif14 in E. coli

目的 :构建 pGEX dif14原核表达载体 ,在大肠杆菌中表达dif14蛋白片段以获得其抗原 .方法 :采用N末端融合谷胱甘肽巯基转移酶 (GST)的 pGEX 4T 1载体 ,根据dif14基因开放读框N端 5 2氨基酸序列设计引物PCR扩增并购建原核表达载体 ,测序证实 .在大肠杆菌DH5α中进行表达 .结果 :测序证实dif14原核表达载体构建正确 ,含 pGEX dif14表达质粒的大肠杆菌经异丙基硫代半乳糖苷 (IPTG)诱导后能够表达约Mr35× 10 3 的融合蛋白 .结论 :成功地构建了原核表达质粒 pGEX dif14 ,并在大肠杆菌得到表达 。

AIM: To construct the expression vector pGEX dif14 and to express the fragment of dif14 protein in E. coli . METHODS: The cDNA coding 52aa of N terminal of dif14 protein was amplified by PCR from dif14 cDNA, inserted into pGEX 4T 1 vector and sequenced. The expression plasmid pGEX dif14 was constructed and expressed in E. coli . RESULTS: The E. coli . containing the expression plasmid pGEX dif14 expressed a 35×10 3 protein after the induction by IPTG. CONCLIUSION: N terminal of dif14 protein has been successfully expressed in E. coli . The study lays a foundation for the functional research of the gene dif14.