大豆半胱氨酸蛋白酶抑制剂基因GmCYS2的功能鉴定

柯丹霞,彭昆鹏,贾妍,曾硕,王英枝,张静怡

Functional Characterization of Soybean Cystatins Gene GmCYS2

Dan-Xia KE,Kun-Peng PENG,Yan JIA,Shuo ZENG,Ying-Zhi WANG,Jing-Yi ZHANG

图3 GmCYS2蛋白的体外表达、抗体检测和蛋白纯化 A: SDS-PAGE分析蛋白在BL21(DE3)菌株中的表达情况; B: Western blot分析, 抗-His抗体检测; C: 纯化蛋白的SDS-PAGE分析。M1: 蛋白marker; M2: Western marker; M3: 蛋白预染marker; PC1: BSA (1 μg); PC2: BSA (2 μg); NC: 未诱导的全细胞裂解液。1: 15℃诱导16 h的全细胞裂解液; 2: 37℃诱导4 h的全细胞裂解液; NC1: 未诱导的细胞裂解液上清液; NC2: 未诱导的细胞裂解液沉淀; 3: 15℃诱导16 h的细胞裂解液上清液; 4: 15℃诱导16 h的细胞裂解液沉淀; 5: 37℃诱导4 h的细胞裂解液上清液; 6: 37℃诱导4 h的细胞裂解液沉淀; 7: 从37℃诱导4 h的细胞裂解液上清液中纯化的蛋白。

Fig. 3 Expression, antibody detection and purification of GmCYS2 protein in vitro A: SDS-PAGE analysis of protein expression in BL21 (DE3) strain; B: Western blot analysis of protein with anti-His antibody; C: SDS-PAGE analysis of purified protein. M1: protein marker; M2: Western marker; M3: protein pre staining marker; PC1: BSA (1 μg); PC2: BSA (2 μg); NC: Non induced whole cell lysate. 1: Whole cell lysate induced under 15°C for 16 h; 2: Whole cell lysate induced under 37°C for 4 h; NC1: Supernatant of non-induced cell lysate; NC2: The precipitation of non-induced cell lysate; 3: Supernatant of cell lysate induced under 15°C for 16 h; 4: The precipitation of cell lysate induced under 15°C for 16 h; 5: Supernatant of cell lysate induced under 37°C for 4 h; 6: The precipitation of cell lysate induced under 37°C for 4 h; 7: Purified protein from supernatant of cell lysate induced under 37°C for 4 h.