Production of transgenic broilers by non-viral vectors via optimizing egg windowing and screening transgenic roosters.

The generation of transgenic chickens is of both biomedical and agricultural significance, and recently chicken transgenesis technology has been greatly advanced. However, major issues still exist in the efficient production of transgenic chickens. This study was designed to optimize the production of enhanced green fluorescence protein (EGFP)-transgenic broilers, including egg windowing at the blunt end (air cell) of egg, and the direct transfection of circulating primordial germ cells by microinjection of the Tol2 plasmid-liposome complex into the early embryonic dorsal aorta.

Functional analysis of the upstream regulatory region of chicken miR-17-92 cluster.

miR-17-92 cluster plays important roles in cell proliferation, differentiation, apoptosis, animal development and tumorigenesis. The transcriptional regulation of miR-17-92 cluster has been extensively studied in mammals, but not in birds. To date, avian miR-17-92 cluster genomic structure has not been fully determined.

[Characterization of chicken PPARγ expression and its impact on adipocyte proliferation and differentiation].

To characterize the chicken PPARγ gene expression and its impact on chicken adipocyte proliferation and differentiation, western blotting approach was conducted to investigate the expression of PPARγ in various chicken tissues and the difference of expression level in abdominal adipose tissues between the NEAU broiler lines divergently selected for abdominal fat content. The expression of PPARγ gene was suppressed in chicken adipocytes using RNAi technology, and the roles of PPARγ gene in the adipocytes proliferation and differentiation were investigated by MTT assay and Oil Red O staining extraction assay, respectively. After PPARγ gene was downregulated, the expression level of other transcript factors and marker genes related to the adipocyte differentiation was detected by Real-time PCR and Western blotting analyses.

[Chicken SREBP1 gene antiserums preparation and its tissue expression characterization].

Aim: The aim of this study was to prepare the antiserums against chicken sterol regulatory element binding protein1 (SREBP1), and to analyze the expression of SREBP1 in chicken tissues.

Methods: The nuclear import sequence of SREBP1 was analyzed using the DNAStar programs to predict its major antigen epitopes, the fragment coding for SREBP1 major antigen epitopes (832-1 302 bp) was amplified by RT-PCR and inserted into pGEX-4T-1 to construct the expression vector pGEX-4T/SREBP1. The recombinant GST/SREBP1 was expressed in E.