Construction and identification of ERβ419 gene silencing recombined vectors of beagle ERβisoforms
10.3969/j.issn.1671.7856.2014.011.003
- VernacularTitle:比格犬 ERβ剪切异构体 ERβ419沉默表达载体的构建及筛选
- Author:
Yi GAN
;
Yanbin ZHAO
;
Fujun CHEN
;
Zhaozeng SUN
;
Lin ZENG
;
Bing LIU
;
Huanmin YANG
;
Zhongming HU
- Publication Type:Journal Article
- Keywords:
Beagles ERβ419;
Stable expression cell lines;
Gene silencing
- From:
Chinese Journal of Comparative Medicine
2014;(11):10-14
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct and identify retroviral-mediated short hairpin RNA ( shRNA ) expression vectors of ERβ419, and explore ERβ419 unknown biological function in beagles in future.Methods To screen out the most effective gene silencing sequence of beagle ERβ419 mRNA using qRT-PCR and Western Blot assays, imitate beagle estrogen target cells.Results qRT-PCR results showed, ERβ419-shRNA1 ( P <0.01 ) and ERβ419-shRNA3 ( P <0.01)differed significantly, Western Blot result as same as qRT-PCR,ERβ419-shRNA3 is the best choice.Conclusion Beagles ERβ419-shRNA3 retrain most effectively target gene repression. It is applied to explore ERβ419 unknown biological function in beagles reproductive system, and to prevent and treat beagles reproductive function diseases.
