四川大学学报(自然科学版)
四川大學學報(自然科學版)
사천대학학보(자연과학판)
JOURNAL OF SICHUAN UNIVERSITY(NATURAL SCIENCE EDITION)
2011年
5期
1197-1203
,共7页
刘昕%吕逢春%杜林方%段伟%刘科
劉昕%呂逢春%杜林方%段偉%劉科
류흔%려봉춘%두림방%단위%류과
酵母%蛋白激酶%表达
酵母%蛋白激酶%錶達
효모%단백격매%표체
Saccharomyces cerevisiae%protein kinase%expression
为了深入研究蛋白激酶Sch9的生物化学特性,在Pichia pastoris酵母KM71H菌种中分泌表达并纯化了重组Sch9蛋白.进一步通过体外磷酸化实验分析了重组Sch9蛋白的蛋白激酶活性.通过重组蛋白质工程的手段初步研究了Sch9蛋白的生物化学和酶学特性.
為瞭深入研究蛋白激酶Sch9的生物化學特性,在Pichia pastoris酵母KM71H菌種中分泌錶達併純化瞭重組Sch9蛋白.進一步通過體外燐痠化實驗分析瞭重組Sch9蛋白的蛋白激酶活性.通過重組蛋白質工程的手段初步研究瞭Sch9蛋白的生物化學和酶學特性.
위료심입연구단백격매Sch9적생물화학특성,재Pichia pastoris효모KM71H균충중분비표체병순화료중조Sch9단백.진일보통과체외린산화실험분석료중조Sch9단백적단백격매활성.통과중조단백질공정적수단초보연구료Sch9단백적생물화학화매학특성.
To facilitate the biochemical characterization of Sch9,the recombinant Sch9 protein was prepared in Pichia pastoris strain KM71H.An N-terminally polyhistidine tagged protein kinase Sch9 was produced and purified via affinity chromatography.After confirming the identity of the recombinant Sch9 protein,was its enzymatic activity further analyzed by an in vitro kinase assay.Our study provides the first insight into the biochemical properties and enzymatic activity of Sch9 via a recombinant protein engineering approach.