Molecular cloning and expression vector construction of bovine TRIM28

The bovine TRIM28 gene was amplified from ovary tissue by using RT-PCR. The TRIM28 gene was inserted into the eukaryotic expression vector pIRES2-EGFP and transfected into bovine fetal fibroblasts by using Lipofectamine 3000. TRIM28 mRNA and protein were detected by fluorescence microscope and weste...

Full description

Saved in:
Bibliographic Details
Published in:Genetics and molecular research 2016-06, Vol.15 (2)
Main Authors: Ma, X, Zhai, Z C, Zhang, M L, Song, B H, Zhu, Y R, Yang, S B, Dong, X Q, Su, L Y, Wang, C F, Ma, H X, Luan, W M
Format: Article
Language:English
Subjects:
Animals
Cattle
Cells, Cultured
Cloning, Molecular
Female
Fibroblasts - metabolism
Genetic Vectors - genetics
Ovary - metabolism
Repressor Proteins - genetics
Repressor Proteins - metabolism
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:The bovine TRIM28 gene was amplified from ovary tissue by using RT-PCR. The TRIM28 gene was inserted into the eukaryotic expression vector pIRES2-EGFP and transfected into bovine fetal fibroblasts by using Lipofectamine 3000. TRIM28 mRNA and protein were detected by fluorescence microscope and western blotting. The results showed that the full length of TRIM28 was cloned and pIRES2-EGFP-TRIM28 was constructed successfully. EGFP expression was observed, and the pIRES2-EGFP-TRIM28 transfected group expressed more TRIM28 protein than that by the pIRES2-EGFP group. The TIMR28 gene has been successfully transferred into bovine fetal fibroblasts.
ISSN:1676-5680
1676-5680
DOI:10.4238/gmr.15028793