我们的网站为什么显示成这样?

可能因为您的浏览器不支持样式,您可以更新您的浏览器到最新版本,以获取对此功能的支持,访问下面的网站,获取关于浏览器的信息:

|本期目录/Table of Contents|

OsRUS2.1酵母双杂交猎物载体的构建及其细胞毒性和自激活作用检测(PDF)

《广西植物》[ISSN:1000-3142/CN:45-1134/Q]

期数:
2013年01期
页码:
76
栏目:
植物生理与分子生物学
出版日期:
2013-01-25

文章信息/Info

Title:
Construction of OsRUS 2.1 yeast twohybrid prey vectors and detection of their toxicity and selfactivated activity
作者:
潘家强1 侯学文12*
1.华南农业大学 生命科学学院 分子植物生理研究室, 广州 510642; 2.华南农业大学 生命科学学院 植物功能基因组与生物技术重点实验室, 广州 510642
Author(s):
PAN JiaQiang1 HOU XueWen12*
1.Lab of Molecular Plant Physiology, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China; 2.Key Laboratory of Plant Functional Genomics and Biotechnology, College of Life Sciences, South China Agricultural University, Guangzhou 510642, China
关键词:
水稻根对UVB敏感基因2.1 猎物载体 酵母双杂交 自激活 毒性检测
Keywords:
rice ROOT UVB SENSITIVE 2.1(OsRUS2.1) prey vector yeast two hybrid selfactivated transcriptional activity toxicity detection
分类号:
Q782
DOI:
10.3969/j.issn.1000-3142.2013.01.014
文献标识码:
A
摘要:
采用PCR技术扩增水稻根UVB敏感基因2.1(ROOT UVB SENSITIVE 2.1,RUS2.1)四个不同片段[OsRUS2.1(11317),OsRUS2.1(1138),OsRUS2.1(139879),OsRUS2.1(8801317)],连接到T载体pMD18TSimple上,测序无误后分别亚克隆到猎物表达载体pGADT7上。结果表明:四个OsRUS2.1基因片段的表达载体构建成功,读码框正确;分别转化这四个猎物表达载体于酵母感受态细胞Y187中,用LacZ、MEL1活性检测法和营养缺陷型培养基SDLeuDO培养法进行自激活检测和毒性检测,结果表明构建的四个OsRUS2.1不同片段的猎物表达载体对酵母菌株Y187均没有转录激活活性和毒害作用,可用于后续研究。
Abstract:
Four fragments of rice(Oryza sativa)ROOT UVB SENSITIVE 2.1(OsRUS2.1),OsRUS2.1(11317),OsRUS2.1(1138),OsRUS2.1(139879),OsRUS2.1(8801317),were amplified by PCR from cloned OsRUS2.1 plasmid,and were ligated with pMD18Tsimple vector,then transformed to E.coli TOP10 competent cell. The positive clones were selected and sequenced. The confirmed fragments were subcloned to prey vector pGADT7. The four constructed pGADT7 prey vectors were further confirmed by enzyme digestion and sequencing. The confirmed 4 types of pGADT7 prey vectors were transformed to Y187 yeast competent cells. The selfactivation and toxicity of the plasmids to host yeast Y187 were detected by LacZ and MEL1 activity assays and culturing in auxotroph medium SDLeu DO. Results showed that the four constructed plasmids had no selftranscriptional activity and not toxicity to yeast strain Y187,and they could be used in the following yeast twohybrid experiments.

参考文献/References

-

备注/Memo

备注/Memo:
-
更新日期/Last Update: 2013-01-25