甘蔗ScWRKY4基因的克隆与表达特性分析

王玲,刘峰,戴明剑,孙婷婷,苏炜华,王春风,张旭,毛花英,苏亚春,阙友雄

Cloning and Expression Characteristic Analysis of ScWRKY4 Gene in Sugarcane

Ling WANG,Feng LIU,Ming-Jian DAI,Ting-Ting SUN,Wei-Hua SU,Chun-Feng WANG,Xu ZHANG,Hua-Ying MAO,Ya-Chun SU,You-Xiong QUE

图7 甘蔗ScWRKY4转录激活活性分析 (A) pGBKT7-ScWRKY4重组载体的酶切验证结果。1: 15000+2000 bp DNA marker; 2: pGBKT7/Nde I; 3: ScWRKY4 ORF PCR产物; 4: pGBKT7-ScWRKY4/Nde I; 5: pGBKT7-ScWRKY4/Nde I+BamH I; 6: 2000 bp ladder DNA marker。(B) ScWRKY4转录激活活性验证。SD/-Trp: 色氨酸营养缺陷型平板培养基; SD/-Trp (+ X-α-Gal): 色氨酸营养缺陷型平板培养基(添加5-溴-4-氯-3-吲哚-α-D-半乳糖苷); SD/-Trp (+ X-α-Gal+AbA): 色氨酸营养缺陷型平板培养基(添加5-溴-4-氯-3-吲哚-α-D-半乳糖苷和金担子素A)。

Fig. 7 Transactivation activity assay of ScWRKY4 in sugarcane (A) The enzyme digestion to identify the insert-integrated vector pGBKT7-ScWRKY4. 1: 15000+2000 bp DNA marker; 2: pGBKT7/Nde I; 3: ScWRKY4 ORF PCR product; 4: pGBKT7-ScWRKY4/Nde I; 5: pGBKT7-ScWRKY4/Nde I+BamH I; 6: 2000 bp ladder DNA marker. (B) The test of transactivation activity assay of ScWRKY4. SD/-Trp: synthetic dropout medium plate; SD/-Trp (+ X-α-Gal): synthetic dropout medium plate without tryptophan (plus 5-bromo-4-chloro-3-indoxyl-α-D-galactopyranoside); SD/-Trp (+ X-α-Gal+ AbA): synthetic dropout medium plate without tryptophan (plus 5-bromo-4-chloro-3-indoxyl-α-D-galactopyranoside and aureobasidin A).