Construction of eukaryotic expression vector encoding rat IMD gene and expression in rat renal tissue via ultrasound-microbubbles
10.3760/cma.j.issn.1008-6315.2011.08.001
- VernacularTitle:中介素真核表达载体的构建及其在超声微泡介导下的肾脏表达
- Author:
Guozhen ZHU
;
Rongshan LI
;
Xi QIAO
;
Xiaoguang HUANG
;
Xiaoqin ZHANG
;
Chen WANG
;
Shan SHAO
;
Bo BAI
- Publication Type:Journal Article
- Keywords:
Intermedin;
Eukaryotic expression vector;
Ultrasound-microbubbles;
Tranfection
- From:
Clinical Medicine of China
2011;27(8):785-788
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct eukaryotic expression vector encoding rat IMD gene and deliver it into rat renal tissue via ultrasound-mircobubbles. Methods IMD gene was inserted into pCDNA3.1 ( + )between Hind Ⅲ and EcoRI enzyme sites. The recombinant plasmid designated as IMD-pCDNA 3.1 wasconfirmed by restrictive enzyme digestion and sequencing. Eighteen male Wistar rats were randomized into 3groups, which were treated with no transfection, empty vector transfection and IMD transfection, respectively, in renal tissue via ultrasound-microbubbles. RT-PCR and Western blotting were applied to detect the expression level of IMD. Results Enzyme- digestion and sequencing data showed that IMD-pCDNA 3.1 was correctly constructed. The differences in ALT, AST, BUN and SCr were not significant; No obvious damage in the glomerular, tubular and interstitial was observed in all the treated groups;Compared with non-transfection group and empty vector-transfection group, IMD mRNA and protein expression in IMD transgenic renal tissue were significantly increased. Conclusion IMD-pCDNA 3.1 expression vector was successfully constructed and well expressed in rat kidney.
