Double-stranded oligomeric nucleotide encoding PEP-1 peptides was synthesized, pro- karyotic expression pET15b-pep-1-p27mt recombinant constructed, E. coli BL21 (DE3)pLysS trans- formed and induced with IPTG to highly...Double-stranded oligomeric nucleotide encoding PEP-1 peptides was synthesized, pro- karyotic expression pET15b-pep-1-p27mt recombinant constructed, E. coli BL21 (DE3)pLysS trans- formed and induced with IPTG to highly express fusion protein PEP-1-P27mt. Fusion protein with an N-terminal His-tag could be purified by Ni2+-resin affinity chromatography and identified by SDS-PAGE and Western blotting. Cultured EC9706 cells treated with PEP-1-P27mt revealed that PEP-1-P27mt was transduced into cells after 15 min and reached maximal intracellular concentra- tions in 2 h. PEP-1-P27mt of 1 μmol/L final concentration could most strongly suppress the growth. It was suggested that PEP-1 can carry P27mt across membrane, which provides a new biological pro- tocol for using cyclin dependent kinase inhibitors p27mt in suppressing the growth of tumor cells.展开更多
本文研究了小麦（Triticum aestivum L.）和大豆（Glycine max L.）的不同光合器官中的PEP羧化酶及其相关的酶类。在这些器官中均含有PEP羧化酶,其中以大豆的荚壳和种皮及小麦的内稃中PEP羧化酶活性最高。不同绿色器官在光下或暗中均能...本文研究了小麦（Triticum aestivum L.）和大豆（Glycine max L.）的不同光合器官中的PEP羧化酶及其相关的酶类。在这些器官中均含有PEP羧化酶,其中以大豆的荚壳和种皮及小麦的内稃中PEP羧化酶活性最高。不同绿色器官在光下或暗中均能固定CO2,但是在黑暗条件下,小麦的内稃和大豆的荚壳及种皮所固定的CO2远高于叶片。此外,参与暗固定的NAD-苹果酸酶和NAD-苹果酸脱氢酶的活性,在这些器官中也最高,说明这些器官中PEP羧化酶的CO2 β-羧化作用主要在于固定呼吸作用所释放的CO2,只有少量的CO2是通过C4光合途径被固定。展开更多
基金a grant from Department of Education of Hubei Province, China (No. Q200524001).
文摘Double-stranded oligomeric nucleotide encoding PEP-1 peptides was synthesized, pro- karyotic expression pET15b-pep-1-p27mt recombinant constructed, E. coli BL21 (DE3)pLysS trans- formed and induced with IPTG to highly express fusion protein PEP-1-P27mt. Fusion protein with an N-terminal His-tag could be purified by Ni2+-resin affinity chromatography and identified by SDS-PAGE and Western blotting. Cultured EC9706 cells treated with PEP-1-P27mt revealed that PEP-1-P27mt was transduced into cells after 15 min and reached maximal intracellular concentra- tions in 2 h. PEP-1-P27mt of 1 μmol/L final concentration could most strongly suppress the growth. It was suggested that PEP-1 can carry P27mt across membrane, which provides a new biological pro- tocol for using cyclin dependent kinase inhibitors p27mt in suppressing the growth of tumor cells.
