%0 Journal Article
%T Overexpression, purification and helicase activity analysis of recombinant human PIF1 protein<br>人类重组PIF1蛋白的表达纯化和解螺旋酶活性的分析
%A Yongqing Gu
%A <br>顾永清
%J 生物工程学报
%D 2009
%I 
%X Pif1 subfamily helicase is conserved from yeast to humans with a lot of cellular functions. In order to elucidate the function of human PIF1 helicase from biochemical level, we cloned human PIF1 gene by PCR from HeLa cell cDNA library. We co-transformed a pMStRNA1 plasmid encoding rare tRNA codons and a plasmid encoding molecular chaperon to greatly enhance the overexpression of human PIF1 protein. Finally we purified full-length PIF1 helicase by column chromatograph carried out at 4°C using fast protein liquid chromatograph (FPLC) system. The human PIF1 protein was purified in enough quantity for detailed biochemical analysis. Biochemical assay showed that PIF1 had ATPase activity and helicase activity. The purification and biochemical properties analysis of human PIF1 helicase will allow us to understand how, at the molecular and mechanistic level, this conserved helicase operate in the cell.
%K PIF1 helicase
%K overexpression and purification
%K ATPase activity
%K helicase activity<br>PIF1解螺旋酶
%K 蛋白表达和纯化
%K ATP酶活性
%K 解螺旋酶活性
%U http://www.alljournals.cn/get_abstract_url.aspx?pcid=90BA3D13E7F3BC869AC96FB3DA594E3FE34FBF7B8BC0E591&jid=A66E90C274451689E69F6F0291467824&aid=B8B7835340A7CCB4CC8C5E476F9D24B7&yid=DE12191FBD62783C&vid=C5154311167311FE&iid=0B39A22176CE99FB&sid=DC330B09A33F1455&eid=9F8C5EF901EA1E7E&journal_id=1000-3061&journal_name=生物工程学报&referenced_num=0&reference_num=13