北柴胡糖基转移酶基因BcUGT3,BcUGT6的表达分析及其原核表达  

Expression analyses of BcUGT3 and BcUGT6,and their in vitro expression in Escherichia coli

作　　者：陶韵文[1,2] 徐洁森[1] 孙晶[1,3] 魏建和[1] 刘娟[2] 隋春[1] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所,北京100193 [2]佳木斯大学药学院,黑龙江佳木斯154007 [3]东北林业大学生命科学院,黑龙江哈尔滨150040

出　　处：《中国中药杂志》2014年第2期185-191,共7页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(81072994);北京市自然科学基金面上项目(5102033);中医药行业科研专项(201107011)

摘　　要：该研究测定了北柴胡中2个糖基转移酶基因BcUGT3和BcUGT6在不同组织中的转录水平及MeJA处理后不定根中的转录水平。BcUGT3在根、叶、花和果中的转录水平无明显差异,但均高于茎中的转录水平。BcUGT6在叶中转录水平最高,在花中最低。以未处理的为对照,BcUGT6在MeJA处理后2 h,8 h,24 h,2 d,4 d的北柴胡不定根中,转录水平均提高近2倍,表明BcUGT6的转录受MeJA影响较小。BcUGT3转录水平随处理时间的延长不断增加,4 d时,达7倍左右。利用载体pET-28a(+),进行了2个基因的原核表达。IPTG诱导后,宿主菌表达出了目的蛋白,并获得了纯化蛋白。为后续开展这2个基因的功能研究奠定了基础。The tissue-specific and MeJA-induced transcriptional levels of BcUGT3 and BcUGT6 in Bupleurum chinense were analyzed in the present study. The transcriptional levels of BcUGT3 in root, leaf, flower and fruit were similar and they all were higher than those in stem. The transcriptional level of BcUGT6 was the highest in leaf and the lowest in flower among in all tested tissues. With non-treated adventitious roots as control, BcUGT6＇s transcriptional levels were elevated to nearly 2 folds for 2 h, 8 h, 24 h, 2 d and 4 d in MeJA-treated adventitious roots of B. chinense. It showed that the transcriptional level of BcUGT6 was slightly affected by MeJA. While, BcUGT3＇s transcriptional levels were gradually elevated, and till 4 d after MeJA treatment, the expression level was about 7 folds than that of non-treated control. Using pET-28a （ ＋ ）, the expressions of two genes was investigated. Induced by IPTG, the target proteins were expressed in E. coli and then purified. All the results obtained in the present study will be helpful for follow- up bio-function analysis of BcUGT3 and BcUGT6.

关 键 词：北柴胡 UGT基因 组织表达 MeJA诱导表达 原核表达 蛋白纯化 

分 类 号：R284[医药卫生—中药学]