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园艺学报 ›› 2006, Vol. 33 ›› Issue (4): 833-835.

• 研究报告 • 上一篇    下一篇

苹果属小金海棠转录因子MxMYB1基因的克隆及其原核表达

曹冬梅1, 2;许雪峰1;韩振海1*   

  1. (1 中国农业大学园艺植物研究所, 北京100094; 2 山西省农业科学院园艺研究所, 山西太原030031)
  • 收稿日期:2005-06-25 修回日期:2005-09-01 出版日期:2006-08-25 发布日期:2006-08-25

Identification and Prokaryotic Expression of Transcription Factor MxMYB1Gene in Malus xiaojinensis

Cao Dongmei1, 2; Xu Xuefeng1;Han Zhenhai1*   

  1. (1 Institute of Horticultural Plants, China Agricultural University, Beijing 100094, China; 2 Institute of Horticulture, Shanxi Agricultural Academy, Taiyuan, Shanxi 030031, China)
  • Received:2005-06-25 Revised:2005-09-01 Online:2006-08-25 Published:2006-08-25

摘要: MxMYB1是苹果属小金海棠MYB类转录因子。MxMYB1含1个重复序列及MYB蛋白特有的氨基酸组成。酶切、PCR扩增及测序分析表明构建的原核表达载体结构正确, 未出现碱基突变及移码现象。1 mmol/L IPTG诱导2 h后, 在预期的蛋白分子量38 kD处出现1条表达加强的蛋白条带, 而未经诱导的转化子没有此蛋白条带。为进一步目的蛋白的纯化和鉴定提供试验基础。

关键词: 转录因子MxMYB1, 原核表达, pET30a-MxMYB1

Abstract: MxMYB1 is a MYB transcription factor from Malus xiaojinensis. MxMYB1 contains a highly conserved DNA binding domain and amino acids existed in MYB proteins particularly. Restriction endonuclease analysis, PCR amplifying and sequencing confirmed that construction was correct and had no base mutant and drift. SDS-PAGE of pET-MxMYB1 proteins induced by 1 mmol/L IPTG showed that there was a strengthened protein band in expectant 38 kD protein Marker, but SDS-PAGE of pET-MxMYB1 proteins uninduced by IPTG had not this band. These results will more provide the foundation for purifying and identifying objective
protein.

Key words: Transcription factor MxMYB1, Prokaryotic expression, pET30a-MxMYB1