hDaxx及其删除突变体在原核细胞中的表达与鉴定

Expression and Identification of hDaxx and Its Deletion Mutants in Prokaryotic Cells

目的 将人Daxx(humanDaxx ,hDaxx)及其 6种删除突变体 (△hDaxx)在原核细胞大肠杆菌BL2 1(E .coli)中表达 ,以便在体外探索hDaxx的生物学活性。方法 用PCR方法从Hela细胞cDNAs文库中扩增全序列hDaxxcDNA ,利用载体pQE3x构建了hDaxx及其△hDaxx质粒 ,通过异丙基 - β -D -硫代半乳糖苷(IPTG)诱导E .coli表达相应蛋白质 ,并将其纯化。结果 IPTG能诱导hDaxx及其△hDaxx在E .coli中表达 ,表达产物纯化率达 85 %以上。结论 hDaxx及其 6种△hDaxx能在E .coli中表达。

Objective:To question the biological activities of human Daxx(hDaxx)in vitro,hDaxx and its 6 deletion mutants(△hDaxx)were induced expression in prokaryotic cells,E.coli BL21.Methods:The prokaryotic expression plasmids of hDaxx and △hDaxx were constructed in pQE3x.Expression of hDaxx and △hDaxx were induced by IPTG.Expression productions were purifed using Ni-NTA agarose.Results:Coomassie strains and Western blot showed that hDaxx and △hDaxx were induced expression by IPTG.Purific rates of expression are more than 85%.Conclusion:hDaxx and its 6 △hDaxx were induced expression in E.coli.