人热休克因子原核表达及表达产物的研究

Studies on Human Heat Shock Factors Expressed in Prokaryotic Cells

利用ＤＮＡ重组技术构建了人ｈｓｐ反式作用因子ＨＳＦ１和ＨＳＦ２的原核表达质粒ｐＥＴ－１１Ｂ／ＨＳＦ１和ｐＥＴ－１１Ｂ／ＨＳＦ２。经ＩＰＴＧ诱导，二者在ＢＬ２１（ＤＥ３）ｐＬｙｓＳ菌株中得到有效表达。利用ｈｓｐ９０β第一内含子ＨＳＥ和合成ＨＳＥ作为探针，对比分析两种表达产物与其特异性顺式作用元件间的结合能力，结果显示，原核表达的ＨＳＦ具有ＨＳＥ结合活性，ＨＳＦ１对ＨＳＥ的亲和力及结合在ＨＳＥ上的分子数明显高于ＨＳＦ２，这种差异可能是两种ＨＳＦ具有不同转录活化功能的分子基础。

uman heat shock factors HSF1 and HSF2 encoding cDNAs were individually cloned intoprokaryotic expression vector pET-11B and were designated as pET-11B/HSF1 and pET-11B/HSF2. Both factors were efficiently expressed in BL21(DE3)pLyS cells under IPTG induction. Heat shock elements(HSE)in the first intron of hsp90β gene was taken as a probe to study thebinding efficiency of,prokaryotic expressed HSF1 and HSF2 in electrophoresis mobility shift as-say, Results showed that both of the HSFs expressed in prokaryotic cells bind to HSE simultane-ously and the binding activity of HSF1 was higher than that of HSF2.