河南农业科学
河南農業科學
하남농업과학
JOURNAL OF HENAN AGRICULTURAL SCIENCES
2014年
10期
26-29
,共4页
杨凤博%王鲜萍%李坤
楊鳳博%王鮮萍%李坤
양봉박%왕선평%리곤
原核表达%纯化%蛋白激酶%脱落酸
原覈錶達%純化%蛋白激酶%脫落痠
원핵표체%순화%단백격매%탈락산
prokaryotic expression%purification%protein kinase%ABA
从哥伦比亚生态型拟南芥(Arabidopsis thaliana Columbia 0)中克隆 SnRK2·6[SNF1(su-crose non-fermenting-1)-related protein kinase 2·6]的完整编码序列(coding sequence,CDS),构建该基因的原核表达载体,将其转化 BL21(DE3),经表达纯化得到 SnRK2·6蛋白。激酶活性分析发现,原核表达纯化的 SnRK2·6有自磷酸化和磷酸化 MBP(myelin basin protein)的活性,为后续试验分析 SnRK2·6的功能奠定基础。
從哥倫比亞生態型擬南芥(Arabidopsis thaliana Columbia 0)中剋隆 SnRK2·6[SNF1(su-crose non-fermenting-1)-related protein kinase 2·6]的完整編碼序列(coding sequence,CDS),構建該基因的原覈錶達載體,將其轉化 BL21(DE3),經錶達純化得到 SnRK2·6蛋白。激酶活性分析髮現,原覈錶達純化的 SnRK2·6有自燐痠化和燐痠化 MBP(myelin basin protein)的活性,為後續試驗分析 SnRK2·6的功能奠定基礎。
종가륜비아생태형의남개(Arabidopsis thaliana Columbia 0)중극륭 SnRK2·6[SNF1(su-crose non-fermenting-1)-related protein kinase 2·6]적완정편마서렬(coding sequence,CDS),구건해기인적원핵표체재체,장기전화 BL21(DE3),경표체순화득도 SnRK2·6단백。격매활성분석발현,원핵표체순화적 SnRK2·6유자린산화화린산화 MBP(myelin basin protein)적활성,위후속시험분석 SnRK2·6적공능전정기출。
The whole coding sequence of SnRK2·6 [SNF1 (sucrose non-fermenting-1 )-related protein kinase 2·6]was cloned from Arabidopsis thaliana ecotype Columbia 0.The prokaryotic expression vector of the gene was constructed and transformed into Escherichia coli strain BL21 (DE3)to obtain SnRK2·6 protein by induction and purification.By activity analysis,the purified SnRK2·6 had both autophosphorylation activity and phosphorylation activity of the myelin basic protein(MBP)substrate.This study lays the foundation for further study of the function of SnRK2·6.