农业科学与技术（英文版）
農業科學與技術（英文版）
농업과학여기술（영문판）
AGRICULTURAL SCIENCE & TECHNOLOGY
2011年
3期
375-378
,共4页
Taq DNA聚合酶%热纯化%pET-32A%BL21(DE3)
Taq DNA聚閤酶%熱純化%pET-32A%BL21(DE3)
Taq DNA취합매%열순화%pET-32A%BL21(DE3)
Taq DNA polyrnerase%Thermal purification%pET-32A%BL21 (DE3)
[目的]提高Taq DNA聚合酶的制备效率.[方法]利用Ni柱亲和色谱纯化载有6xHis标记的Taq DNA聚合酶,并重组载体,利用Taq DNA聚合酶的耐热特性,对粗提液75℃处理1h,之后通过PCR试验验证制备酶液的活力.[结果]所获重组的pET-32A-Taq能够在BL21(DE3)宿主菌中高效表达并可通过热变性去除杂蛋白,获得了活力远高于购买的Taq DNA聚合酶的酶制剂.[结论]使用热纯化法制备的Taq DNA聚合酶工艺简单,成本较低,能满足常规大量PCR实验要求.
[目的]提高Taq DNA聚閤酶的製備效率.[方法]利用Ni柱親和色譜純化載有6xHis標記的Taq DNA聚閤酶,併重組載體,利用Taq DNA聚閤酶的耐熱特性,對粗提液75℃處理1h,之後通過PCR試驗驗證製備酶液的活力.[結果]所穫重組的pET-32A-Taq能夠在BL21(DE3)宿主菌中高效錶達併可通過熱變性去除雜蛋白,穫得瞭活力遠高于購買的Taq DNA聚閤酶的酶製劑.[結論]使用熱純化法製備的Taq DNA聚閤酶工藝簡單,成本較低,能滿足常規大量PCR實驗要求.
[목적]제고Taq DNA취합매적제비효솔.[방법]이용Ni주친화색보순화재유6xHis표기적Taq DNA취합매,병중조재체,이용Taq DNA취합매적내열특성,대조제액75℃처리1h,지후통과PCR시험험증제비매액적활력.[결과]소획중조적pET-32A-Taq능구재BL21(DE3)숙주균중고효표체병가통과열변성거제잡단백,획득료활력원고우구매적Taq DNA취합매적매제제.[결론]사용열순화법제비적Taq DNA취합매공예간단,성본교저,능만족상규대량PCR실험요구.
[Objective] The paper wes to improve the preparation efficacy of Taq DNA polymerase. [Method] Ni column was used to purify Taq DNA polymerase carrying with 6xHis tag, and recombined vector. Using the thermal-resistant characteristics of Taq DNA polymerase, the crude extract was treated at 75 ℃ for 1h, and the activity of prepared enzyme solution was verified by PCR test. [Result] The recombinant pET-32A-Taq could highly express in BL21 (DE3) host bacteria and remove hybrid protein by thermal denaturation. The enzyme preparation with the activity further higher than purchased Taq DNA polymerase was obtained. [Conclusion] Taq DNA polymerase prepared by thermal purification method is simple with Iow cost, and can meet the needs of a large number of conventional PCR amplification.
